AN ABSTRACT OF THE DISSERTATION OF Doolalai Sethajintanin for the degree of Doctor of Philosophy in Toxicology presented on September 2, 2005. Title: Passive Sampling Devices as Biological Surrogates for Evaluating Seasonal Bioavailability of Hydrophobic Organic Contaminants in Surface Water. Redacted for Privacy Abstract approved Kim A. Anderson The seasonal distribution of bioavailable organochiorine contaminants in surface water and the potential environmental factors influencing their bioavailability were evaluated. The study was carried at the lower Willamette River at Portland Harbor, Oregon where surface water runoff varied according to season. Bioavailable water concentrations of DDTs and PCBs were determined using a polyethylene membrane containing passive sampling device (PSD), known as semipermeable membrane device (SPMD). Our findings indicated that the influence of river seasonality on the bioavailable distributions of organochiorine contaminants was compound- and site-specific. Bioavailable DDTs concentrations were strongly affected by the local historic use of DDTs and seasonal changes in river conditions. The dominance of bioavailable p,p'-DDD and large DDD/DDE ratios observed during low flow condition in summer suggest redistribution of p,p'-DDD into the water column and conditions favoring reductive dechlorination of p,p'-DDT to p,p'-DDD. In contrast, bioavailable dieldrin and PCB concentrations were significantly increased during high flow condition in fall, especially during episodic rainstorm events. While a discernable seasonal pattern for PCBs was observed along the 18-mile stretch study area, the seasonal pattern of dieldrin was only apparent at the sampling site downstream of an agricultural creek with historical use of dieldrin. The increase in bioavailable PCB concentrations and daily loads cOincident with high precipitation and sewer overflows in fall suggested a significant contribution of PCBs from precipitation input and urban storm water discharges to the surface water. Seasonal bioavailable concentrations of organochiorine compounds exceeded the national and the Oregon water quality criteria revealing the significance of considering realistic seasonal and site-specific influences on bioavailable organochiorine distributions when performing risk assessments. In addition, we developed the triolein-free polyethylene lay flat tubing (LFT) as an alternative in situ PSD. The LFT proved reliable and had the same benefits as SPMD, but was simpler, inexpensive and lacked interference from the triolein impurities. The LFT tended to accumulate compounds with high log K0 faster than SPMD. LFT sampling rates were estimated and modeled for 33 target analytes, including PAHs, PCBs, and organochiorine pesticides. The successful determination of field derived data illustrates the effectiveness and reliability of LIT for environmental monitoring. © Copyright by Doolalai Sethajintanin September 2, 2005 All Rights Reserved Passive Sampling Devices as Biological Surrogates for Evaluating Seasonal Bioavailability of Hydrophobic Organic Contaminants in Surface Water by Doolalai Sethajintanin A DISSERTATION submitted to Oregon State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Presented September 2, 2005 Commencement June 2006 Doctor of Philosophy dissertation of Doolalai Sethajintanin presented on September 2, 2005. APPROVED: Redacted for Privacy Major Professor, representing Toxicology Redacted for Privacy Head of the Department of Environmental and Molecular Toxicology Redacted for Privacy Dean of th- radiiate School I understand that my dissertation will become part of the permanent collection of Oregon State University Libraries. My signature below authorizes release of my dissertation to any reader upon request. Redacted for Privacy Doolalai Sethajintanin, Author ACKNOWLEDGMENTS Many people have contributed, in various ways, to this accomplishment, and it is not feasible in limited space to mention them all. I would like to express my sincere appreciation to my major professor, Dr. Kim A. Anderson, for providing guidance, encouragement, partial financial support and many opportunities for regional and national conferences. Her advice and generosity are appreciated. I acknowledge my graduate committee, Drs. Lawrence R. Curtis, Jeffrey J. Jenkins, Staci L. Simonich and Daniel D. Rockey, for contributing suggestions and comments to this study. Special thanks go to Dr. Jennifer A. Field for reviewing and contributing valuable comments to Chapter 2. This study was partially funded by the Society of Environmental Toxicology and Chemistry (SETAC) Early Career for Applied Ecological Research Award sponsored by the American Chemistry Council awarded to Dr. Kim A. Anderson. The Oregon Department of Environmental Quality for the McCormick & Baxter superfund site project 2002-2003 and the Oregon Health Science University pilot project for the NIEHSIEPA Superfund Basic Research Grant provided partial financial support. I am deeply indebted to the Royal Thai Government Scholarship for financial support. The Food and Drug Administration, Ministry of Public Health, Thailand, is acknowledged for providing me the opportunity to pursue my master and doctor degrees. I thank all of my teachers for contributing to my knowledge. I am grateful for field assistance by Robert Grove of Forest and Rangeland Ecosystem Science Center, Corvallis, Oregon. It is a pleasure to acknowledge the field and laboratory assistance from my colleagues at the Anderson Lab group; Eugene Johnson, Bobby Loper, Solyssa Visalli, Gregory Sower, Jennifer Basile, Roger Adams, Angela Perez, Oraphin Krissanakriangkrai, Lucas Quarles, and Dr. Wendy Hiliwalker. I am especially grateful to Eugene Johnson and Bobby Loper for their wonderful kindness. Special thanks go to the Anderson Lab group, all of my friends at Oregon State University, my officemates (Kimberly Hageman and Junga Lee) and my little Thai community who made my days in Corvallis memorable. I would like to thank my Thai friends in Thailand and the US, especially Yuda Chongpison and Arusa Chaovanalikit, for supporting and encouraging me emotionally. Last but not least, I would like to thank the most important people in my life, my mom and dad (Prajoub and Chinta Sethajintanin), for always being there and loving support although physically thousands of miles away. Without their loves and inspiration this accomplishment would not have been possible. CONTRIBUTION OF AUTHORS This study was conducted with the contributions of the following co-authors. Dr. Kim A. Anderson provided guidance on all aspects of this dissertation. Eugene R. Johnson participated in the preparation of triolein-free low density polyethylene membrane in Chapter 3 and the chemical analysis work in Chapter 4. TABLE OF CONTENTS Page CHAPTER 1 INTRODUCTION 1 Background 5 Bioavailability 5 Significance of spatial and seasonal aspects on chemical distribution 7 Passive sampling technique as surrogate for evaluating bioavailability 8 Organochiorine compounds in aquatic system ..12 Study area: the lower Willamette River at Portland Harbor, Oregon 15 Hypotheses and objectives .17 Literature Cited 18 CHAPTER 2 EVALUATION OF SEASONAL BIOAVAILABILITY OF ORGANOCHLORINE PESTICIDES AND PCBS AT A SUPERFUND SITE USING PASSIVE SAMPLING DEVICE 23 Abstract 24 Introduction 25 Materials and Methods Results and Discussion .. .28 .33 River conditions and water chemistry .33 Distributions of bioavailable DDT concentrations and Potential sources 36 Dominance of bioavailable p, p'-DDD in surface water TABLE OF CONTENTS (Continued) Page Evidence of seasonality in bioavailable DDT concentrations 41 Potential risk of seasonal bioavailable DDTs on aquatic organisms 45 Distribution of bioavailable polychlorinated biphenyls (PCBs) 48 Seasonal changes in bioavailable PCB distribution 50 Bioavailable PCB composition in surface water 50 PCBs-associated risks to aquatic community and human health 54 Seasonal changes in bioavailable dieldrin distribution Literature Cited CHAPTER 3 FIELD TRIAL OF TRIOLEIN-FREE LOW DENSITY POLYETHYLENE MEMBRANE AS AN IN SITU PASSIVE WATER SAMPLER . .55 57 .63 Abstract 64 Introduction 65 Materials and Methods 68 Results and Discussion 73 Evaluation of the spiking method of permeability/performance Reference compounds (PRCs) in LFT 73 Demonstration of effectiveness and reliability of LFT for field deployment .75 Literature Cited 92 TABLE OF CONTENTS (Continued) Page CHAPTER 4 ENVIRONMENTAL STRESSES AND FISH DEFORMITIES IN THE WILLAMETTE RIVER: COMPARISON OF PERSISTENT BIOACCUMULATIVE TOXICANTS .96 Abstract 97 Introduction 98 Materials and Methods .100 Results and Discussion .103 Water chemistry characterization 103 Bioavailable polychiorinated biphenyls (PCBs) .. 106 p, p'-DDT and its metabolites 107 Dieldrin 108 Literature Cited .109 CHAPTER 5 CONCLUSIONS ..111 BIBLIOGRAPHY .116 APPENDIX A Estimated river concentrations of bioavailable p,p'-DDT and its metabolites, dieldrin, and PCBs (sum of 25 congeners) at the Willamette River during summer and fall of 2001 to 2004. River concentrations were estimated from SPMD concentrations. Deployments of SPMDs were typically 7-to 21-days long 126 APPENDIX B Sewer overflow points in Portland area .. . .131 APPENDIX C Equations used to estimate molecular diffusivity in water (D) .. . .132 APPENDIX D Predicted values for LFT sampling rates using different methods 133 APPENDIX E Environmental stresses and skeletal deformities in fish from the Willamette River, Oregon 134 LIST OF FIGURES Figure Figure 1.1 Figure 1.2 Page Schematic representation of bioavailability as modified from Dickson et al. 1994 and Wells and Lanno 2001 5 A simple four-phase model describing the bioavailabiiity and the partitioning approach to the distribution of organic contaminant in water-sediment systems .6 Figure 1.3 Generalized uptake profile for a passive sampling device 10 Figure 1.4 The three compartment model for contaminant accumulation by SPMD as proposed by Gale 1998 Figure 2.1 Figure 2.2 Figure 2.3 Figure 2.4 Figure 2.6 Figure 2.7 Figure 2.8 ..11 The lower Willamette River at Portland, Oregon, showing locations of the sampling sites (RM = river mile) .29 Average concentrations of bioavailable DDTs in surface water at the lower Willamette River at Portland Harbor, Oregon, using passive sampling device (n=186) 38 Distributions of bioavailable p, p'-DDD: p, p'-DDE ratios in surface water at the Willamette River, Oregon during the periods of low flow in summer, and high flow in fall from 2001 to 2004 (n=186) 40 Estimated concentrations of bioavailable p, p'-DDT and its metabolites in surface water at River Mile 7West of the lower Willamette River at Portland Harbor, Oregon 42 Spatial and seasonal distribution of bioavailable p, p' -DDT and its metabolite water concentration estimates compared to the national recommendation water criteria and Oregon criteria 47 Average concentration estimates of bioavailable PCB (pgIL) in water at the Willamette River, Oregon according to river season Average percentage contribution of PCB congeners in surface water at the lower Willamette River during the periods of low flow in summer and high flow in fall in 2001 to 2004 .49 51 LIST OF FIGURES (Continued.) Figure Figure 2.9 Figure 2.10 Figure 2.11 Figure 3.1 Figure 3.2 Figure 3.3 Figure 3.4 Figure 3.5 Figure 3.6 Page Average ratios of PCB homolog to bioavailable PCBs (sum of 25 individual PCB congeners) in surface water at the lower Willamette River . ..52 Spatial and seasonal distribution of bioavailable PCB water concentration estimates (sum of 25 individual PCB congeners) compared to the national recommendation water criteria and Oregon criteria 55 Seasonal distribution of bioavailable dieldrin in surface water at the lower Willamette River at Portland Harbor, Oregon during the period of low flow in summer and high flow in fall of 2001-2004 56 Relationship of contaminant concentrations in the triolein-free polyethylene lay flat tubing (LIT) and in semipermeable membrane device (SPMD) 81 Comparison of spatial distributions of selected contaminants accumulated in the triolein-free polyethylene lay flat tubing and in semipermeable membrane device (SPMD) 83 Percent comparison of PCBs and PAHs (average ± lsd) in the triolein-free polyethylene lay flat tubing (LFT) and semipermeable membrane device (SPMD) .84 Average ratios of concentration of contaminants accumulated in the triolein-free polyethylene lay flat tubing (LFT) and in semipermeable membrane device (SPMD) 84 Sampling rate estimates (RL) of a 540 cm2 triolein-free polyethylene lay flat tubing (LIT) as a function of log K0, molecular volume, molar volume, and minimal molecular two-dimension Sampling rate estimates of a 540 cm2 triolein-free polyethylene lay flat tubing (LIT) as a function of the product of aqueous diffusitivity, log K0, and minimal molecular two-dimension . 88 90 LIST OF FIGURES (Continued.) Figure Figure 4.1 Figure 4.2 Page The Willamette Basin showing the general location and course of the Willamette River and primary study locations 99 The mean concentration distribution of bioavailable p, p' -DDT, p, p'-DDD, and p, p'-DDE at Newberg and Corvallis in 2002 and 2003.. ..107 LIST OF TABLES Table Table 1.1 Pace List of target PCBs and organochiorine pesticides and their physicochemical properties at 20-25 °C 14 Summary of river conditions and water chemistry parameters at Portland Harbor during SPMD deployment 34 Summary results of total organic carbon (TOC), dissolved organic carbon (DOC), total suspended solids (TSS), and total dissolved solids (TDS) at the Willamette River, Oregon during SPMD deployment .35 Table 3.1 Selected physicochemical properties of target analytes .69 Table 3.2 Percent recoveries of permeability/performance reference compounds using simple spiking method in a small volume of Table 2.1 Table 2.2 Table 3.3 Table 3.4 toluerie .74 Percent recoveries and clearance rates of permeability/performance reference compounds in the field exposed polyethylene lay flat tubing (LFT) and semipermeable membrane device (SPMD) under different field condition 78 Contaminant sampling rates for triolein-free polyethylene lay flat tubing (LFT) estimated from field exposure to contaminated water at the lower Willamette River at Portland Harbor, Oregon for 2ldays 87 Table 4.1 Spatial and temporal variations of water quality parameters at Newberg (River Mile 44-47) versus Corvallis (River Mile 135) sampling sites on the Willamette River, Oregon .104 Table 4.2 Mean concentrations (pg/L) of bioavailable organochlorines estimated from concentrations accumulated in semipermeable membrane devices (SPMDs) exposed for 21 days at the Willamette River at Newberg and Corvallis (2 sampling sites per location) in 2002 and 2003 .. 106 PASSIVE SAMPLING DEVICES AS BIOLOGICAL SURROGATES FOR EVALUATING SEASONAL BIOAVAILABILITY OF HYDROPHOBIC ORGANIC CONTAMINANTS IN SURFACE WATER CHAPTER 1 INTRODUCTION Understanding the distribution of chemicals in the environment is required to assess exposure to humans and biota. The environmental transport of hydrophobic organic contaminants such as polychlorinated biphenyls (PCBs), p,p'-DDT (l,l-bis(4-chlorophenyl)2,2,2-trichioroethane) and polycyclic aromatic hydrocarbons (PAHs) are dependent on their distribution properties among various environmental phases. These distribution characteristics are modeled by phase-distribution equilibrium constants such as water solubility, octanol-water partition coefficient (K0), sorption coefficient (Kd or K0), vapor pressure (Vp), and airwater partition coefficient (KH)(]). In aqueous environments, these contaminants exist in diverse states in surface water. They can be freely dissolved in surface water or associate with dissolved or particulate organic matter depending on their chemical and physical properties (2). All of these forms can be transported by water or distributed throughout surface water and their concentrations may exert adverse effects on aquatic organisms and human health through multiple sources and pathways with variable and often poorly understood factors. The freely dissolved form of hydrophobic organic contaminants is considered environmentally relevant to their bioavailability, toxicity, mobility and degradation processes (3-5). It is the freely dissolved form of the contaminant which is transported across biological membranes of aquatic organisms and potentially exerts toxic effects (2, 4, 5). A reduction in the freely dissolved concentration of the contaminant refers directly to reduced bioavailability. However, there is no clear distinction between the processes that control the distribution of chemical contaminants in an environment and those that directly affect bioavailability (6). Spatial and temporal factors, including seasonal changes in physical, 2 chemical and biological aspects of the aquatic ecosystem, may modify contaminant bioavailability (i.e., aqueous activity and fugacity) and change the concentration available for uptake by aquatic organisms. The potential uptake of toxic contaminants is affected by changes of either the concentrations of the contaminant in the organism's immediate environment or the environmental bioavailability of the contaminant (6). Bioavailability is a key to deteiiiiining the nature of the exposure and the toxicity of a chemical in the environment. However, it is a problematic parameter to measure. Bioavailability, by definition, is determined by target concentrations in organisms (5). Any chemical estimates of bioavailabiiity must be correlated with biological measures of bioavailability or toxicological bioavailability (5). Recent developments in passive sampling devices provide an alternative to understand chemical bioavailability (7). Instead of measuring the total concentration in an environmental medium, the passive sampling technique measures the concentration in a reference phase which can be brought into equilibrium with the medium (7). The availability of a chemical is measured based on the chemical potential which is logarithmically related to its fugacity and linearly related to its freely dissolved concentration in a particular medium (7). As correlated with biological concentrations, chemical estimates using passive sampling devices can predict toxicity and can be used as a screening tool in the first tier of ecological risk assessment. Identifying and measuring the chemical forms that are actually the bioavailable forms is necessary to better understand the physical, chemical, and biological mechanisms that control contaminant fates and effects in the aquatic environment. Advanced understanding of bioavailable chemical distribution is required to predict accurately the fate and effects of environmental contaminants and this knowledge can have substantial benefits for exposure assessments and water-quality regulations. Despite the importance of environmental bioavailability, very few studies have actually evaluated the distribution of chemical bioavailability and the influencing factors in the field under different and often more variable geochemical or physiological conditions than in the laboratory. As a part of an assessment of the bioavailability of a chemical contaminant, the ultimate goals of this dissertation were to understand the seasonal distribution of bioavailable 3 hydrophobic contaminants in surface water and to evaluate the potential environmental factors influencing their bioavailability. In addition, further development of a passive sampling device to measure bioavailable hydrophobic contaminants and field trial were conducted and its efficiency was compared to a widely used passive sampling device. The dissertation chapters are described below. This chapter (Chapter 1) contains a general introduction providing background information needed for the study. It contains the basic concept of bioavailability, passive sampling technique, and general background of the target analytes and study area. The dissertation hypotheses and objectives are present at the end of the chapter. Chapter 2 presents the field study of distribution and seasonal variation of bioavailable organochiorines in surface water using passive sampling techniques. The study was carried out at the lower Willamette River at Portland Harbor, Oregon where surface water runoff is determined by climatic influence and varies according to season (8). The study area (River Mile 1 to 18) included the Portland Harbor superfund site (River Mile 3.5 to 9.5) and the McCormick Baxter superfund site (River Mile 7 East) (9). The potential environmental factors influencing their bioavailable concentration distributions are discussed. To screen the potential impacts associated with seasonal changes on aquatic organisms, the bioavailable concentrations are compared to the national recommended water quality criteria (10) and the Oregon water quality criteria (11). Chapter 3 presents the further development of a passive sampling device. The field trial of triolein-free low density polyethylene membrane was studied. Its efficiency was compared to the widely used passive sampling device. The sampling rates and uptake model for a wide range of hydrophobic contaminants are proposed. Chapter 4 is part of a collaborative study of "Environmental stresses and skeleton deformities in fish from the Willamette River, Oregon" (12). This chapter focuses only on the distribution of bioavailable PCBs and organochiorine pesticides along specific sections of the Willamette River (Newberg Pool, River Mile 44 to 47), where elevated frequencies of skeletal deformities in fish were detected, relative to those in the upper river (Corvallis, River 4 Mile 135). The complete collaborative work has been published in Environmental Science and Technology. Volume 39, No. 10, 2005, page 3495-3506, "Environmental stresses and skeletal deformities injlshfrom the Willainette River, Oregon" (12), see Appendix E. Chapter 5 summarizes the overall conclusion of this dissertation. The direction for future work is also provided. 5 Background Bioavailability. Bioavailability of a contaminant is specific to the exposure matrix, duration of exposure, route of entry, and receptor. The concept of bioavailability consists of three major components (13) as described in Figure 1.1. Environmental availability represents the portion of the total material present in a compartment or compartments of the environment which actually involves a particular process or group of processes and is subject to modifying influences (13). However, the total amount of chemical potentially available is not necessarily involved in a given process. Bioavailability is a special case of environmental availability when organisms are involved in the process as a target. Bioavailability may be described as the portion of the total quantity or concentration of a chemical in the environment or a portion of it that is potentiaLly available for biological action, such as uptake by an aquatic organism (14). Toxicological bioavailability is the fraction of the dose absorbed by the organism which reaches the target sites in the organism or effective intercellular concentration in the organism (13). Environmental availability Environmental bioavailability Toxicological bioavailability Total chemical Sorbedl sequestered Freely dissolved Absoi ion Distribution Metabolism Excretion Site of toxic action Bioaccurnulation Media Exposure Organism Figure 1.1 Schematic representation of bioavailability as modified from Dickson et al. 1994 (13) and Wells and Lanno 200 1(15). 6 Water column w, org ingestion or ,' excretion ," ingestion or excretion rnembrane' transport K11 or BCF sorption! desorption mass transport advective or diffusive flux qj K0 "bou .ar layer" sorption! dcsorption V SV. ti K.or sorption! desorption ingestion or excretion s, sq C sorptiol1! A desorption oenihrane transport '.. doe KdOC KB or BCFY ,/ s, doe mass transport resuspension "boun.ar Ia er" sedimentation ;4 ingestion or excretion C s,org Sediiient Figure 1.2 A simple four-phase model describing the bioavailability and the partitioning approach to the distribution of organic contaminant in water-sediment systems. The freely dissolved fractions of the contaminants (Cw,aq and Cs, aq) are the most bioavailable. The contribution from bound fractions (C,55, CW,d00, C,55 and C5, doe) may become important for certain organism via ingestion as part of the food supply. The first subscript indicates the environmental compartment; w = water, s = sediment. The second subscript refers to the phase within that compartment; aq = aqueous (freely dissolved), org = organism, doc = dissolved organic carbon, ss = suspended solids. (modified from Suffet et al. 1994 (2)) In aquatic systems, if the uptake through ingestion of food is excluded, the freely dissolved fractions of organic contaminants are the most bioavailable (5). Organic chemicals that are freely dissolved in the water, and not bound to or associated with organic matter, are subject to diffusion transport across aquatic organism's membrane from the external aqueous (2). Figure 1.2 illustrates a simple four-phase model describing the bioavailability and the partitioning approach to the distribution of organic contaminant in water-sediment systems. 7 The aqueous concentration at equilibrium is related to the distribution coefficient (K) of the various phases, assuming the interactions of the contaminant concentrations are at equilibrium in the water column and sediment. Partitioning to sediments, and dissolved or particulate organic matter decreases freely dissolved concentration of the contaminant and thus the bioavailability of contaminants to biota (2, 5). Many factors can influence the environmental availability including the environmental bioavailability by altering the environmental distribution of the chemicals and this would consequently alter bioconcentration and bioaccumulation (2, 6, 14). Physical factors include temperature, advection, and sedimentation and resuspension. Physicochemical factors include dissolution, desorption, diffusion, and degradation. Biological factors include diagenesis, organic matter loading and bioturbation. However, there is no clear distinction between the processes that control the distribution of chemical contaminants in an environment and those that directly affect bioavailability (6). Significance of spatial and seasonal aspects on chemical distribution. A consideration of the spatial and seasonal aspects is essential to chemical exposure in ecological risk assessment (16). These variables affect the concentrations available for uptake from surrounding media by organisms. Several studies have indicated bioaccumulation of PCBs and DDTs in fish and lower trophic level biota seasonally changed according to concentrations of PCBs and DDTs in surrounding water and sediment (17, 18). As well, a significant spatial and seasonal pattern in exposure modeling for human health risk estimates for recreational fish consumers has been addressed (18). Consideration of spatial and temporal characteristics of contaminant bioaccumulation in fish reduced risk estimates as much as one order of magnitude lower than those obtained without the spatial and temporal consideration. In addition to improving our understanding of the chemical fate and transport in the environment, understanding and consideration of spatial and seasonal distribution of contaminants can reduce uncertainty associated with chemical exposure and provide a quantitative expression of the confidence in risk estimates. Consideration of spatial and seasonal variation can help support the decision-making process for risk assessment and risk management of contaminated water and sediment. 8 Spatial factors include sources of chemical contaminants, location of sensitive biological resources, routes of exposure, and factors that may modify contaminant mobility and availability (i.e., changing composition of sediment, abundance of microorganisms involved in contaminant biodegradation) (19). Temporal factors include seasonal changes in physical, chemical, or biological aspects of ecosystem that may impact the potential for exposure (19). The influence of seasonal variation on contaminant concentration and load may vary across a wide spectrum of the watershed characteristic. Foster et al. (20) reported increased water movement during storm flow enhanced PCB, organochiorine pesticides, and PAH riverine transport in both dissolved and particulate phase at the Susquenhanna River Basin, Maryland, USA. Soderstrom et al. (21) observed increased concentrations of DDTs and changes in DDD: DDE ratios (DDT metabolites) after snowmelt due to resuspension of surface sediment in a eutrophic and an oligotrophic lake. By contrast, PCB concentrations in the Eman River, Sweden were inversely related to river discharge due to dilution processes (22). In addition, seasonal variation in physical or chemical parameters (i.e., temperature or pH) can modify the bioavailability of contaminants and consequently change the nature of exposure (19). Although the spatial and seasonal distributions of chemical contaminants such as PCBs and organochlorine pesticides have been widely studied, significant information gaps exist for their seasonal bioavailability. It is the freely dissolved fraction, not particulate fraction or dissolved fraction associated with dissolved organic matter, that is most bioavailable and relevant to the internal concentrations of the target organisms (5). Because PCBs and organochiorine pesticides are a problem of global concern, such information gaps must be addressed to promote a more complete understanding of their bioavailability characteristics that may be influenced by spatial and seasonal factors. Passive sampling techniques as surrogates for evaluating bioavailability. Bioavailability is the key to determining the nature of the exposure and the toxicity of a chemical in the environment. Any chemical estimates of bioavailability must be correlated with biological measures of bioavailability or toxicological bioavailability (5). Measuring the actual dose or toxicant concentration at the target site is ideal for biologically effective dose and toxicity assessment (5). Alternatively, whole body internal concentrations are 9 sufficiently good approximations of target-site concentrations for toxic chemicals (5). However, measuring bioavailable chemicals in the exposed organisms in a field conditions sometimes fails to accurately reflect environmental contaminant concentrations because of residue metabolism and depuration and the effects of environmental stressors on organism health (23). Recent developments in passive sampling devices provide an alternative to understand chemical bioavailability (7, 24, 25). These new developments in sampling technique fit well into chemical uptake and bioconcentration / bioaccumulation in certain aquatic organisms (5, 15, 25-27). Instead of measuring the total concentration in an environmental medium, the passive sampling technique measures the concentration in a reference phase which can be brought into equilibrium with the medium (7 28). The availability of a chemical is measured based on the chemical potential which is logarithmically related to its fugacity and linearly related to its freely dissolved concentration in a particular medium (7, 28). Figure 1.3 illustrates three uptake phases for a passive sampling device, which includes the generalized uptake profile according to Equation 1 - a first-order kinetic model (7): Csampier (t) = Cmedium kL. (1- e2t) (1) where Csampier (t) is the concentration of the contaminant in the sampler as a function of time, t, Cmedium is the concentration in the environmental medium, and k1 and k 2 are the uptake rate and the elimination rate constants, respectively. 10 equiJibrium C O.5 Cu C U) 0 C 0 C.) 0 T112 time Figure 1.3 Generalized uptake profile for a passive sampling device. The passive sampling technique generally operates in three regimes: linear or kinetic, curvilinear or intermediate, and equilibrium. Low density polyethylene membranes, typically filled with triolein, have been successfully deployed as passive sampling devices designed to accumulate nonpolar and moderately polar organic contaminants from water, sediments, and air (15, 27, 29, 30). This device is known as semipermeable membrane device (SPMD) introduced by Huckins et al. (25). It Consists of polyethylene lay flat tubing containing a thin film of a neutral lipid, 95% pure triolein (1,2,3-tri[cis-9-octadecenoyl]glycerol) (25). The diameters of the transient cavities on polyethylene membrane range up to 10 A, which effectively preclude sampling of any contaminant molecules associated with dissolved organic matter or particulates (31). It has been proposed that the SPMD mimics key mechanisms of bioconcentration including diffusing through biomembranes and partitioning between organism lipid and the sunounding medium, but without metabolism (31). For example, uptake of organochiorine contaminants was similar in SPMD, fish, and mussels (25-27). Determining dissolvedlbioavailable concentrations with this technique would be a better measure to assess bioconcentration / bioaccumulation factors in aquatic organisms as compared with filtered water methodology, leading to a better prediction of environmental risk and risk assessment. The three compartment model describing the mechanism for SPMD accumulation has been established (28). The model consists of three compartments; water, polyethylene membrane, and triolein, and three mass transfer step as described in Figure 1.4 by Gale (28). 11 Figure 1.4 shows a dissolved chemical is transported to the aqueous diffusion film via turbulent-diffusive transport through the aqueous film and partioning into polyethylene membrane, and diffusive transport through the polyethylene film, and partitioning between it and the triolein. It is noted that polyethylene as a chemical reservoir in SPMD is accounted for by the three compartment model and it is worth the effort to further develop it as an alternative passive sampling device as present in Chapter 3. A Ditlusional SJow Steps c' W-ter PoteriiiI Biotirn Ple h1orie Trioer Steps Turbulent transport (rnxinç) 0 ii Pnpor lCl t eylow 1 n Ti [C) n [CL Tr B Water Potyethytene Trioein Figure 1.4 The three compartment model for contaminant accumulation by SPMD as proposed by Gale, 1998 (28). A) Proposed passive sampling steps for SPMD accumulation. B) The three compartment model consisting of water, polyethylene and triolein. Rate constants for input into the water (k0),accurnulative mass transfer from water to polyethylene (k), from polyethylene to triolein (kpT), and the corresponding clearance rate constants for mass transfer from water to polyethylene (k) and from polyethylene to triolein (kT) are shown with their respective anows. (Figure is adapted from Gale, 1998) 12 The linear uptake model (Figure 1.3) is desired for the integrative sampling approach for SPMD application (23). Huckins et al. (23) have derived the first order kinetic model (Equation 1) to estimate bioavailable water concentrations from SPMD concentrations in the linear uptake phase and can be rewritten as = -SPMD Fl] SPMD. Ilc . t -1 (2) where C'SPMD is the concentration of the bioavailable contaminant in the SPMD, MSPMD is the mass of SPMD in grams R is the sampling rate in liters per day of a standard 1-g triolein SPMD, and t is the exposure time is days. Using Equation (2) and available Rç values (26, 32, 33), the bioavailable water concentrations of hydrophobic contaminants can be estimated. For exposure conditions of low to moderate flow/turbulence, SPMD uptake is under membrane control for compounds with log K0 < 4.4 and under aqueous boundary layer control for compounds with log K0 4.4 (23). Thus the SPMD uptake of chemicals with log K0 4.4, which include most PCB congeners, organochiorine pesticides, and PAHs, are sensitive to site hydrodynamics. SPMD uptake rates are also affected by temperature and biofouling (34). Booij et al. (24) and Huckins et al. (34) have developed the use of peiiiieability / performance reference compound (PRC) to provide an overall correction factor for variations in SPMD uptake rates under field condition. Organochuorine compounds in aquatic system. Polychlorinated biphenyls (PCBs) and organochiorine pesticides including p,p'-DDT and its metabolites (p,p'-DDD and p,p'DDE) and dieldrin are industrial and agricultural compounds. These compounds have environmental significance because of their stability, toxicity, and tendency to accumulate in lipids of organisms (35-37). Their production and use have been banned since 1970s. The current major source of these compounds releases to air, water, and soil is from the cycling of their residues remaining in the environment from one medium to another. Atmospheric transport also plays an important role in long-range transport of organochiorines from their initial source (38). 13 Organochiorine environmental transport varies depending on their chemical and physical properties. Table 1.1 lists physicochemical properties of target PCBs and organochiorine pesticides in this dissertation. Organochiorines exist in diverse states in surface water. They can be freely dissolved in surface water or associated with dissolved or particulate matter depending on their chemical and physical properties (35-37). All of these forms can be transported by water or distributed throughout surface water. Their major transport is mainly via waterborne particles but in a river containing small amount of particles, their major transport can occur in the dissolved phase (35-37). In addition to ingestion of PCBs or DDTs contaminated in the aquatic food chain, aquatic organisms (e.g. fish, mollusk) directly uptake bioavailable form of these contaminants from surrounding water into their body by passive diffusion through semipermeable membranes such as gill, lining of the mouth, or gastrointestinal tract (35-37, 39). After uptake, organochiorine contaminants are transported to different compartments of the body including sites of action, metabolism, storage and excretion. The toxic effects of a contaminant can differ between species, strains, sex and age groups (35, 36, 40). DDT is highly toxic to aquatic organisms (35). Early developmental stages of fish and aquatic invertebrate are more susceptible than adults to DDT (35). The acute toxicity of p,p'-DDT to both vertebrates and invertebrates is attributed mainly to action upon axonal Na* channels (41). Apart from the action upon Na channels, it has been reported that p,p'-DDT can act upon the K channels and inhibit certain ATPase (4]). In fish, cellular respiration may be the main toxic target of DDT due to the inhibition of ATPase which can affect osmoregulation (35, 41). The effects of DDT on a range of physiological functions and behavioral development in fish have been reported (35). It has been reported that DDT can cause skeleton deformities by impairing developmental processes and bone formation in certain fish species (42). Pathologic changes by DDT are also observed in the liver and reproductive organs of laboratory animals (35). 14 Table 1.1 List of target PCBs and organochiorine pesticides and their physicochemical properties at 20-25 °C chemical - Trichiorobiphenyls PCB 37 - Tetrachiorobiphenyls PCB 44 PCB49 water solubility (mg/L)a Log K0 a 3,4,4' 1.5 x 102 2,2',3,5' 2,2',4,5' 2,2',5,5' 2,3,4,4' chemical structure PCB 52 PCB 60 PCB 74 2,4,4',S PCB 77 3,3',4,4' - Pentachiorobiphenyls PCB 87 2,2',3,4,5' PCB 99 2,2',4,4',5 PCB 101 2,2,4,5,5' PCB 105 2,3,3',4,4' PCB 114 2,3,4,4',5 PCB 118 2,3',4,4',5 PCB 126 3,3',4,4',5 - Hexachiorobiphenyls PCB 128 2,2',3,3',4,4' PCB 138 2,2',3,4,4',5' PCB 153 2,2',4,4'5,5' PCB 156 2,3,3',4,4',5 PCB 166 2,3,4,4',5,6 PCB 169 3,3',4,4',5,5' Heptachiorobiphenyls PCB 170 2,2',3,3',4,4',5 PCB 180 2,2',3,4,4',5,5' PCB 183 2,2',3,4,4',5',6 PCB 187 2,2',3,4',5,5',6 PCB 189 2,3,3',4,4',5,5' Organochlorine pesticides sorption coefficient, log K0 a vapor pressure 5.9 4.8 4.5 x i0 1.0 x 10_i 6.0 6.1 6.1 6.3 6.7 6.5 4.7 5.7 3.9 5.7 6.4 x i0 1.6x102 3.0 x 10.2 4.0 x 102 NA 1.0 x i0 4.0 x i0 2 1.1 x 10 1.0 x 102 NA NA NA NA 6.5 6.6 6.4 6.0 6.7 7.1 6.9 6.0 x 10 7.0 1.5 x i0 1.0 x i0 NA NA 5.1 x 10 6.7 6.9 5.0 x i0 3.1 x i0 7.2 7.0 7.6 NA 4.4 (Pa a 7.4x103 2.0 x i0 2.2 x 10 NA 2.0 x i0 5.7 5.7 5.7 NA NA NA NA 2.3 x 10 5.3 5.9 5.9 NA NA 6.6 3.4 x i0 5.0 x i0 7.0 x i0 1.5 x i0 3.5 x l0 NA NA NA NA NA NA 5.4x i0 7.2 7.2 7.2 7.7 5.6 5.8 NA 5.5 NA 5.5 x i0 5.7 6.3 2.5 x 10 p,p'-DDD 2.0 x 102 6.1 5.0 1.3 x 10 p,p'-DDE 1 x 10' 6.0 4.7 8.7 x 10 dieldrin 2 x 10' 3.5 4.1 4.0 x iO4 p,p'-DDT . NA 4.0 x i0 NA 7.1 1.3 x 10 3.1 x i0 NA 9.4 x i0 NA NA - not available a Sources: Mackay et al. (43), Augustijn-Beckers et al. (44), Petty et al.(45), and Eisler and Belisle (46). 15 Dieldrin is highly toxic to aquatic crustaceans and fish (36). Several studies have revealed that dieldrin toxicity increases with increasing temperature (36). It has been reported that dieldrin can produce adverse enzymatic and hormona' change in fish that lead to impaired reproductive ability (36). Different life stages of fish have been found to have different susceptibility to dieldrin. Eggs were resistant and juvenile stages were less susceptible than adults (36). Many studies have reported PCB toxicity in both in vitro and in vivo studies (40). The toxicity of individual PCBs is related to the molecular structure. The most potent congeners are those that contain non-ortho or mono-ortho chlorine substituents, so called coplanar PCBs, which have a similar structure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. These dioxin-like PC]3s act upon aryl hydrocarbon (Ah) receptors, with consequent induction of cytochrome P450 1A1/2 and Ah-receptor mediated toxicity (i.e. immunotoxicity, disruption of multiples endocrine pathways, developmental toxicity, reproductive toxicity, carcinogenicity, tumor promotion, hepatotoxicity) (40). Non-planar PCBs elicit toxic responses including neurobehavioral, neurotoxic, carcinogenic and endocrine changes by acting through multiple unrelated mechanisms (47). Although the toxicology of PCBs has been studied extensively, the degree of toxicity and nature of the effects remain highly debatable. The complexity of PCB contamination with the possibility of interactive effects between different congeners andlor with other persistent contaminants have made ecological effects of PCBs difficult to prove (41). Recently, the influence of PCBs on the endocrine system has been a subject of particular environmental interest. Aquatic communities may be susceptible to endocrine disruption due to their association with PCB contaminated sediments and surface water. Many studies have suggested PCBs adversely affected gonadal development, vitellogenin expression, thyroid function and sexual differentiation in aquatic organisms (37, 40, 46, 48). It has been suggested that the decline in certain fish populations may be associated with developmental and reproductive impairment caused by PCBs (48). Study area: the lower Willamette River at Portland Harbor, Oregon. The Willamette River in western Oregon is one of only 14 American Heritage Designated Rivers 16 (49). The river has the thirteen the largest stream flow in the United States and yields more runoff per square mile than any other large river in the US (50). The Willamette River flows approximately 187 miles northward through Portland, Oregon's largest metropolitan area, before joining the Columbia River. After this joining, the Columbia River flows an additional 100 miles westward to the Pacific Ocean. The Willamette River provides a significant migratory corridor, nursery habitat and adult forage for runs of salmon, and nearly 50 species of fish have been identified in the river (51). Recreational and sport fishing are extremely popular throughout the lower Willamette basin. Most development along the Willamette River has been located within the Portland Harbor. Portland Harbor is heavily industrialized, and contains a multitude of facilities and both private and municipal waste water outfalls. The harbor generally has a low sediment transport capacity (8). Surface water runoffs are determined by climate influence and vary according to season (8). The presence of PCBs, pesticides, PAHs, dioxins/furans, arsenic, cadmium, chromium, copper, lead, mercury, zinc, organotins, pentachiorophenol, and solvents has been demonstrated in Portland Harbor sediment within a six-mile stretch from the southern tip of Sauvie Island to Swan Island (RM 3.5 RM 9.5)(8). These contaminants may have entered the river via spillage during product shipping and handling, direct disposal or discharge, accidental spills, contaminated ground water discharge, surface water runoff, stormwater discharge, or contaminated soil erosion (52). These contaminants can pose risks to people, fish and other wildlife and the detected levels were high enough to place Portland Harbor on the Federal National Priority List (NPL, commonly known as superfund) (52). The NPL is the US Environmental Protection Agency (EPA)'s list of the national most contaminated hazardous waste sites which may pose dangers to public health and the environment and are therefore targeted for cleanup (9). Portland Harbor became a superfund site on December 1, 2000 (9). The area covers the six-mile stretch between the southern tip of Sauvie Island to Swan Island (RM 3.5 RM 9.5) and contains McCormick and Baxter Creosoting Co. (Portland plant) superfund site. Our previous study found PCB and DDT residues accumulated in three recreational fish species from the Portland Harbor exceeded the US EPA fish advisory's screening values and potentially posed adverse health effects to the recreational and subsistence fishers (53, 17 54). Bioaccumulation of PCBs and DDTs in fish is related to the concentrations of PCBs and DDTs in surrounding water and sediment and can change seasonally (17 18). Consequently, the seasonal pattern in exposure modeling for environmental and human health risk assessment can affect the accuracy of risk estimates (18). As surface water runoffs within the harbor are determined by climate influence and vary according to season (8), PCB and DDT river concentrations, particularly the bioavailable concentrations may change seasonally and consequently change the nature of exposure. This can affect both aquatic organisms and human healTh. The river characteristics make Portland Harbor an excellent candidate for studying the seasonal bioavailability of PCBs and organochiorine pesticides in surface water. Hypotheses and objectives Hypothesis 1. The water concentrations of bioavailable organochiorines including PCBs, p,p'-DDT and its metabolites (p,p'-DDD and p,p'-DDE), and dieldrin change seasonally (Chapter 2). Objectives - To determine the bioavailable concentrations of organochlorines in surface water using passive sampling device - To evaluate the spatial influence on the distribution of bioavailable organochiorines in surface water - To evaluate the seasonal distribution of bioavailable organochiorines in surface water and to assess the potential factors influencing their seasonal concentrations Hypothesis 2. Seasonal changes affect the nature of exposure to organochlorines in surface water by aquatic organisms and local fishers who consume fish in the study area (Chapter 2). Objective - To screen the potential impacts associated with seasonal changes on aquatic organisms and local fish consumers by comparing seasonal concentrations of bioavailable organochiorines to the national recommended water quality criteria (10) and the Oregon water quality criteria (11) 18 Hypothesis 3. The solvent- free low density polyethylene membrane lay flat tubing (LFT) is as efficient as the widely used passive sampling device in sampling bioavailable hydrophobic organic contaminants in surface water (Chapter 3). Objectives - To evaluate the use of LFT as an alternative device for monitoring bioavailable hydrophobic organic contaminants in surface water - To compare the efficiency of LFT with the semipeimeable membrane device (SPMD) Literature cited Schwarzenbach RP, Gschwend PM, Imboden DM. Environmental organic chemistry. New York, New York: John Wiley & Sons, Inc., 1993. Suffet III, Jafvert CT, Kukkonen J, Servos MR, Spacie A, Williams LL, Noblet JA. Synopsis of discussion session: influences of particulate and dissolved material on the bioavailability of organic compounds. In: Bioavailability:physical, chemical, and biological interactions (Benson WH, ed). Boca Raton: Lewis Publishers, 1994. Walker CH, Hopkin SP, Sibly RM, Peakall DB. 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Huckins JN, Petty JD, Lebo JA, Almeida FV, Booij K, Alvarez DA, Cranor WL, Clark RC, Mogensen BB. Development of the permeability/performance reference compound approach for in situ calibration of semipermeable membrane devices. Environmental Science and Technology 36:85-91(2002). WHO/IPCS. DDT and its derivatives - environmental aspects. Geneva: World Health Organization, 1989. WHO/IPCS. Aidrin and dieldrin. Geneva: World Health Organization, 1989. WHO/IIPCS. Polychiorinated biphenyls and terphenyls (second edition). Geneva: World Health Organization, 1993. Wania F, Mackay D. Tracking the distribution of persistent organic pollutants. Environmental Science and Technology 30: 390A-396a( 1996). McKim JM. Physiological and biochemical mechanisms that regulate the accumulation and toxicity of environmental chemicals in fish. In: Bioavailability: physical, chemical, and biological interactions (Benson WH, ed). Boca Raton, FL: Lewis Publishers, 1994; 179-201. Safe SH. 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United States Environmental Protection Agency. American Heritage Designated Rivers. Last update December 3, 2003 [cited May 25, 2005]. Available from http://www.epa. gov/river/98rivers/ Uhrich MA, Wentz DA. Environmental setting of the Willamette basin, Oregon WaterResources Investigations Report 97-4082-A. Portland: US Department of the Interior US Geological Survey, 1999. Altman B, Henson CM, Waite JR. Summary of information on aquatic biota and their habitats in the Willamette River basin, Oregon, through 1995 Water-Resources Investigations Report 97-4023. Portland, Oregon: US Geological Survey, 1997. State of Oregon Department of Environemental Quality. Portland Harbor superfund site. Last update December, 2004 [cited May 10, 2005]. Available from http://www.deq. state.or.us/nwr/portlandharbor/ph .htm Sethajintanin D. Bioaccumulation profiles of chemical contaminants in fish from the lower Willamette River, Portland Harbor superfund site and human health risk assessment. In: Environmental and Molecular Toxicology. Corvallis: Oregon State University, 2003; 119. Sethajintanin D, Johnson ER, Loper BR, Anderson KA. Bioaccumulation profiles of chemical contaminants in fish from the lower Willamette River, Portland Harbor, Oregon. Archives of Environmental Contamination and Toxicology 46:114-123(2004). 23 CHAPTER 2 EVALUATION OF SEASONAL BIOAVAILABILITY OF ORGANOCHLORINE PESTICIDES AND PCBs AT A SUPERFUND SITE USING PASSIVE SAMPLING DEVICES Sethajintanin, D.1 and Anderson, K.A 1,2 Department of Environmental and Molecular Toxicology 2 Food Safety and Environmental Stewardship Program, Oregon State University, Corvallis, Oregon 97331 24 Abstract Although the spatial and seasonal distribution of polychiorinated biphenyls (PCBs) and organochiorine pesticides (i.e., DDT, dieldrin) have been widely studied, significant information gaps exist for their bioavailability. Because PCBs and organochiorine pesticides are a problem of global concern, such information gaps must be addressed to promote a more complete understanding of their bioavailability characteristics that may influence spatial and seasonal factors, The present study focused on the spatial and seasonal distribution of bioavailable organochiorine pesticides and PCBs within the surface water of a contaminated harbor. Passive sampling devices were intensively deployed just outside the shipping channel adjacent to various land uses on the lower Willamette River, Oregon including Portland Harbor superfund site in summer and fall during 2001-2004. An increase of estimated bioavailable DDTs (sum of p, p'-DDT, p, p'-DDD, and p,p'-DDE) concentrations was strongly affected by the local historic productions of DDTs and seasonal changes in river conditions. The dominance of bioavailable p, p'-DDD and high DDD/DDE ratios observed during periods of low flow in summer suggested direct inputs of p, p' -DDD and conditions favoring reductive dechlorination of p, p'-DDT top, p'-DDD. The estimated bioavailable concentrations and daily loads of PCJ3s and dieldrin increased during periods of high precipitation, high river flow in fall, especially during episodic rainstorms. The similarity of PCB congener profiles and PCB homolog ratios at the industrial area and at the urban/residential areas suggested the local sources of PCBs at the industrial area were important enough to significantly increase the concentrations of bioavailable PCBs in surface water but not so substantial as to change composition relative to surface water inputs from upstream. Bioavailable concentrations organochlorine compounds seasonally exceeded the national and the Oregon water quality criteria. These seasonal exceedances suggest the potential impacts associated with seasonal changes of bioavailable organochiorine distributions in surface waters and the significances of considering realistic seasonal and site- specific conditions in risk assessment and water quality management. Keywords Willamette River bioavailability organochiorines contaminants seasonal influence 25 Introduction The freely dissolved form of hydrophobic organic contaminants is considered environmentally relevant to their bioavailability, toxicity, mobility and degradation processes (1-3). It is the freely dissolved form of the contaminant which is transported across biological membranes of aquatic organisms and potentially exerts toxic effects (2-4). A reduction in the freely dissolved concentration of the contaminant refers directly to reduced bioavailability, and vice versa. However, there is no clear distinction between the process that controls the distribution of chemical contaminants in an environment and those that directly affect bioavailability (5). Spatial and temporal factors including seasonal changes in physical, chemical and biological aspects of the aquatic ecosystem may modify contaminant bioavailability and this changes concentration available for uptake from water by aquatic organisms. The potential uptake of toxic contaminants is affected by changes in either the concentrations of the contaminant in the organism's immediate environment or the environmental bioavailability of the contaminant (5). Despite the importance of environmental bioavailability, very few studies have actually evaluated the distribution of chemical bioavailability and the influencing factors in the field under different and often more variable geochemical or physiological conditions than in the laboratory. A consideration of the spatial and seasonal aspects is essential to chemical exposure in ecological risk assessment (6). These variables affect the concentrations available for uptake from surrounding media by organisms. Several studies have indicated bioaccumulation of polychiorinated biphenyls (PCBs) and DDTs (1,1-bis(4-chlorophenyl)2,2,2-trichioroethane and its metabolites) in fish and lower trophic level biota seasonally changed according to concentrations of PCI3s and DDTs in surrounding water and sediment (7, 8). As well, a significant spatial and seasonal pattern in exposure modeling for human health risk estimates for recreational fish consumers has been addressed (8). Consideration of spatial and temporal characteristics of contaminant bioaccumulation in fish reduced risk estimates as much as 1 order of magnitude lower than those obtained without the spatial and temporal consideration. In addition to improving our understanding of chemical fate and transport in the environment, understanding and consideration of spatial and seasonal distribution of contaminants can reduce uncertainty associated with chemical exposure and 26 provide a quantitative expression of the confidence in risk estimates. Consideration of spatial and seasonal variation can help support the decision-making process for risk assessment and risk management of contaminated water and sediment. Spatial factors include sources of chemical contaminants, location of sensitive biological resources, routes of exposure, and factors that may modify contaminant mobility and availability (9). Temporal factors includes seasonal changes in physical, chemical, or biological aspects of ecosystem that may be a potential for exposure (9). The influence of seasonal variation on contaminant concentration and load may vary across a wide spectrum of watershed characteristic. Foster et al. (10) reported increased water movement during storm flow enhanced transport of PCBs, organochiorine pesticides, and polycyclic aromatic hydrocarbons (PAHs) in both dissolved and particulate phase at the Susquenhanna River Basin, Maryland, USA. Soderstrom et al. (11) observed increased concentrations of DDTs and changes in DDD: DDE ratios after snowmelt due to resuspension of surface sediment in a eutrophic and an oligotrophic lake. By contrast, PCB concentrations in the Eman River, Sweden were inversely related to river discharge due to dilution process (12). In addition, seasonal variation in physical or chemical parameters (i.e., temperature or pH) can modify the bioavailability of contaminants and consequently change the nature of exposure (9). Although the spatial and seasonal distributions of chemical contaminants such as PCBs and DDTs have been widely studied, significant information gaps exist for their seasonal bioavailability. Because PCBs and organochiorine pesticides are a problem of global concern, such information gaps must be addressed to promote a more complete understanding of their bioavailability characteristics that may be influenced by spatial and seasonal factors. The Willamette River in western Oregon is one of only 14 American Heritage Designated Rivers (13). The river has the thirteenth largest stream flow in the US and yields more runoff per square mile than any other large river in the US (14). Recreational and sport fishing are extremely popular throughout the lower Willamette basin. The lower end of the Willamette River, Portland Harbor, is heavily industrialized and contains a multitude of facilities and both private and municipal waste water outfalls. The lower Willamette River at Portland Harbor is deep, slow moving, and tidally influenced (15). Due to elevated 27 concentrations of PCBs, organochiorine pesticides, dioxins/furans, PAHs, and heavy metals in the harbor sediment, Portland Harbor has been placed in the Federal National Priority List (NPL), commonly known as superfund (16). Our previous study found PCB and DDT residues accumulated in three recreational fish species from the Portland Harbor exceeded the US EPA fish advisory's screening values and potentially posed adverse health effects to the recreational and subsistence fishers (17, 18). Bioaccumulation of PCBs and DDTs in fish is related to the concentrations of PCBs and DDTs in surrounding water and sediment and can change seasonally (7 8). As surface water runoffs within the harbor are determined by climate influence and vary according to season (15), PCB and DDT river concentrations, particularly the bioavailable concentrations may change seasonally and consequently change the nature of exposure. The consequence can affect both aquatic organisms and human health. The river characteristics make Portland Harbor an ideal study area for evaluating seasonal bioavailability of PCBs and organochiorine pesticides in surface water. The ultimate goals of this study were to understand the seasonal distribution of bioavailable PCBs, p,p'-DDT and its derivatives, and dieldrin in surface water and to evaluate the potential environmental factors influencing their bioavailability. The passive sampling technique using semipermeable membrane device (SPMD) (19, 20) was chosen to get a time-integrated measure of their bioavailable fraction. The SPMD consists of polyethylene lay flat tubing containing a thin film of a neutral lipid, 95% pure triolein (l,2,3-tri[cis-9-octadecenoyl]glycerol) (21). It has been proposed that SPMD mimics key mechanisms of bioconcentration including diffusing through biomembranes and partitioning between organism lipid and the surrounding medium , but without metabolism (19). In this work, the influences of seasonal and episodic events coupled with in situ water chemistry on their bioavailable transport were intensively investigated. The potential environmental factors influencing their bioavailability were discussed. To screen the potential impacts associated with seasonal changes on aquatic organisms, the bioavailable concentrations were compared to the national recommended water quality criteria (22) and the Oregon water quality criteria (23). 28 Materials and Methods Materials and chemicals. Standard SPMDs were purchased from Environmental Sampling Technologies (EST, St. Joseph, MO). Standards of organochiorine pesticides (purities 98.5%) and PCBs (purities 99%) were from Chem Service, Inc. (West Chester, PA) and AccuStandard (New Haven, CT), respectively. All solvents used were pesticide or Optima® grade from Fisher Scientific (Fairlawn, NJ). Certified reference material of PCB congeners and organochlorine pesticides in cod liver oil (SRM 1588a) was purchased from the National Institute of Standards and Technology (NIST, Gaithersburg, MD). SRM 1588a was used for method validity and accuracy purposes for the determination of PCBs and organochlorine pesticides in complex lipophilic matrices such as triolein. Selection of 25 target PCB congeners were based on their toxicity, frequency of occurrence, and abundance in environmental matrices (24). Target PCB congeners included dioxin-like congeners (tetra-C13; PCB 77, penta-CB; PCB 105, PCB 114, PCB 118, PCB 126; hexa-CB; PCB 156, PCB 169; hepta.-CB; PCB 189) and non dioxin-like congeners (tn- CB; PCB 37, tetra-CB; PCB 44, PCB 49, PCB 52, PCB 60, PCB 74, penta-CB; PCB 87, PCB 99, PCB 101, hexa-CB; PCB 128, PCB 138, PCB 153, PCB 166, hepta-CB; PCB 170, PCB 180, PCB 183, PCB 187). Target organochiorine pesticides were p,p'-DDT, p,p'-DDD, p,p'DDE, and dieldrin. Study area. The sampling areas were located at the lower Willamette River at Portland Harbor from River Mile 1 to 18 and contained the Portland Harbor superfund site (River Mile 3.5 to 9.5) and the McCoiiiiick and Baxter Creosoting Co. (Portland plant) superfund site (River Mile 7 East). The sampling sites were outside the main shipping channel and often near stream outlets of various upstream land uses. The eleven sampling sites (Figure 1) were at River Mile 1 East (Columbia Slough; industrial area), River Mile 3.5 West (Sauvie Island at Multnomah Channel; industrial and urban area), River Mile 3.5 East (opposite to Multnomah Channel; industrial area) , River Mile 7 West (Railroad Bridge; industrial area), River Mile 7 East (McCormick and Baxter superfund site), River Mile 8 East (Swan Island; industrial area), River Mile 12 East (under Steel Bridge! Interstate 5 Highway; downtown/urban area), River Mile 13 West (Hawthorne Bridge; downtown/urban area), 29 River Mile 15 East (Ross Island: a sand and gravel operation! undeveloped area), River Mile 17 East (golf course, park), and River Mile 18 West (Mouth of Johnson Creek; urban/agricultural creek, residential area). The Willamette Basin \wasngton Portland ci, 0 0 0 (ti 0 0 0 --Swan Island V Oregn I "aipPortIand - aliforni. 'Nevada 12 RM 13" I 1 L 0 0 4 15 rRM l5'c, RM1 RM18 sampling site 2 -' -' Ross Island - 't tim fN 10km Figure 2.1 The lower Willamette River at Portland, Oregon, showing locations of the sampling sites (RM = river mile) Sources of precipitation and stream flow data. Precipitation and stream flow data were needed to characterize the river conditions during sample collection. Precipitation data were obtained from the National Weather Service Forecast Office - Portland, OR (25). The station was located at the Portland International Airport which was 9 miles northeast of downtown Portland. Stream flow data recorded at River Mile 12.8 were obtained from the US Geological Survey (26). Stream flow data were also used to convert estimated surface water concentrations using SPMD into loading estimates. A daily mass load was calculated by multiplying the estimated river concentration using SPMD by the average daily flow rate through the river. 30 Sample collection. Five individual SPMDs were loaded in a flow-through stainless steel cage. Each cage was submerged approximately 10 ft from the river bottom and was suspended with "anchor-cable-cage-cable-float" arrangement. The river depth at sampling locations ranged from 2-50 ft. At the sampling sites at RM 7 East where the river was generally less than 10 ft. deep, the samplers were suspended below water surface but above the sediment. SPMDs were deployed for 7-21 days. Water temperature and other water chemistry parameters (dissolved oxygen, specific conductivity, oxidation-reduction potential, pH, NH4tN, and NO3 -N concentrations) were measured during deployment and retrieval using an YSI 6920 SONDE (Yellow Springs, OH). The average temperature ranged from 9 to 22 °C. Water samples were also collected for the analysis of total organic carbon (TOC), dissolved organic carbon (DOC), total suspended solids (TSS), and total dissolved solids (TDS). On retrieval, SPMDs were cleaned by gently rubbing with gloved hands in on-site water, then rinsed in iN HC1 (Trace metal grade, Fisher Chemical, Fairlawn, NJ), 18 M2cm water, acetone (Pesticide grade, Fisher Scientific, Fairlawn, NJ), and isopropanol (Pesticide grade, Fisher Scientific, Fairlawn, NJ), respectively. Cleaned SPMDs were kept in clean glass amber jars and transported on ice-packs. Samples were stored at -20 °C until analysis. Sample extraction and chemical analysis. SPMD extraction and cleanup were based on established protocols and only modified slightly as necessary (19, 27). SPMDs were dialyzed in hexanes, 400 mL for 5 SPMDs (Pesticide grade, Fisher Scientific, Fairlawn, NJ) for 18 hr, followed by a second dialysis with fresh hexanes for 6 hr. The combined dialysates were concentrated by rotary evaporation and a TurboVap® LV evaporator (Zymark®, Hopkinton, MA). The samples were cleaned and fractionated using gel permeation chromatography (Waters® gel permeation chromatography cleanup system; Water® 515 HPLC pump, 717 Plus autosampler, 2487 dual X absorbance detector, and fraction collector II, Milford, MA) with dichloromethane (Optima®, Fisher Scientific, Fairlawn, NJ) as the mobile phase at the flow rate of 5 mL/min. Appropriate fractions were determined by analyzing standards and fortified samples. The appropriate fraction (14-20 mm) was collected and split into two equal sub-fractions: 1) PCB and organochlorine pesticides for this study 2) PAHs for another study. PCB and organochlorine pesticide fraction was subjected to volume 31 reduction using a TurboVap® LV evaporator and solvent exchanged into iso-octane (Pesticide grade, Fisher Scientific, Fairlawn, NJ) with a final volume of 1 mL. PCB congeners and OC pesticides were deteiiiiined using GC-ECD (Agilent Technologies 6890N Network GC system, Palo Alto, CA) dual capillary columns (db-xlb and db-17 with length 30 m, i.d. 0.25 mm, and film thickness 0.25 Itm, J&W Scientific Inc., Agilent Technologies, Palo Alto, CA) Idual detectors. Injector and detector temperature were at 250 °C, and 350 °C, respectively. The GC system was operated with helium carrier gas and nitrogen makeup gas. The oven temperature program was as follows: 100 °C (1 mm) and increased at 1.2 OCminl to 265 °C (2 mm). Field duplicates and blanks (trip blank, field blanks, and field extraction blanks) accompanying the deployed passive sampling devices during deployment, retrieval, and transportation to the laboratory were included in every batch. These field blanks and laboratory control blanks (i.e., procedural blanks, laboratory SPMD controls, fortified samples, and standard reference material) represented 30 to 40% of a sample set and were processed and analyzed exactly as the deployed samples. The method detection limits (MDL) were determined as three times the heights of coincident peaks observed for each compound in the laboratory SPMD control. For those analytes having no coincident peak, the MDLs were set at a value equivalent to the lowest standard in the respective calibration curve. Because contaminant uptake rates by SPMDs can change due to changes in temperature, flow velocity of the surrounding water and buildup of periphyton on the membrane surface, an in situ measurement to account for these field variables is needed (28, 29). PCB 8, PCB 82 and endrin were used as permeability/performance reference compounds (PRCs) to measure the overall variations in SPMD uptake rates under field conditions. PRCs are analytically non-interfering compounds with moderate to relatively high fugacity from SPMDs and have physico-chemical properties similar to the target analytes, which are added to the SPMDs prior to sealing the membrane tubes (29). Because both the uptake and the dissipation of organic chemicals are controlled by the same molecular processes, in situ dissipation rates of PRCs would allow for estimating in situ SPMD uptake rates under 32 various field conditions (28, 29). The PRC approach has been well described by Booij et al. (28) and Huckins et al.(29). None of the target analytes were identified in field blanks and laboratory control blanks. The average percent recoveries of fortified samples were as follows: PCBs 69 ± 43%, p,p'-DDT 71 ± 27%, p,p'-DDD 80 ± 21%, p,p'-DDE 66 ±16%, and dieldrin 73 ± 18%. The average percent recoveries in certified reference material were as follows: PCBs 94 ± 26%, p,p'-DDT 98 ± 18%, p,p'-DDD 105 ±17%, p,p'-DDE 76 ± 16% and deildrin 89 ± 6%. Data analysis. The basic theory and mathematical models required for estimation of analyte water concentrations from the concentrations in the SPMD have been described by Huckins et al. (20). The average percent recoveries of PRCs in the deployed SPMDs indicated the linear uptake model could be assumed for all target analytes in this study. Differences in exposure temperature were corrected by using established SPMD sampling rates (R) for PCBs and organochiorine pesticides at multiple temperatures (30-32). The small variations of PRC dissipation rates among sampling sites and under different field seasons suggested the effects of membrane biofouling and flow velocity-turbulence at the membrane surface were considered negligible. Estimates of water concentrations (Cm) from SPMD concentrations were calculated by the following equation (20), C = CSPMD 'MSPMD 'R, where C SPMD is the concentration of the individual analyte in the SPMD, MSPMD is the mass of SPMD in grams, MSPMD is the sampling rate of a standard 1-g triolein SPMD, and t is the time in days. Data interpretation was performed using SPSS® Version 10.0.1(SPSS Inc., 19891999), Sigma Plot 2002 for Windows Version 8.0 (SPSS Inc., 1986-200 1), and Microsoft® Office Excel 2003 (Microsoft Corporation, 1985-2003). Standard descriptive statistics, two sample t-test and linear regression techniques were used to examine seasonal bioavailable 33 organochiorine concentrations and influence variables. Statistical analyses were considered significant atp 0.05. Estimates of bioavailable concentrations in surface waters were compared to the national recommended water quality criteria (22) and the Oregon water quality criteria (23). These guidelines were used as a screening tool to assess potential impacts associated with seasonal or episodic changes of contaminant distributions in surface waters. Fresh water aquatic life criteria are to protect the aquatic communities from toxic effects resulting from chronic exposure to toxic contaminants in water (22). This criterion concentration is an estimate of the highest concentration in surface water to which an aquatic community can be exposed without resulting in an unacceptable effect. Human health water quality criteria are to protect local fishers based on a 106 increased lifetime cancer risk from consumption of fish and shellfish from the concerning areas(22). Results and Discussions River conditions and water chemistry. The lower Willamette River at Portland Harbor could be typically characterized by seasonally defined conditions. We categorized the river conditions according to the average river flow, total precipitation, and average temperature during sample deployments. Over the sample deployment interval from 2001 to 2004, the river was categorized into low flow/low precipitation in summer (average river flow < 10,000 ft3 Is, total precipitation 0-1 in., average temperature 20 °C) and high flow/high precipitation in fall (average river flow> 10,000 ft3 /s, total precipitation> 1 in., average temperature <20 °C). In addition, any intermittent precipitation or high river flow that occurred during sample deployment was considered as an episodic event in the present study. 34 Table 2.1 Summary of river conditions and water chemistry parameters at Portland Harbor during SPMD deployment. river total water flow rain temp. (ft3/s) (in.) (°C) 08/02/01- 7000 0 22 7.4 08/16/01 (170) (0.57) 09/13/01- 7200 09/20/01 (130) 10/16/01- 13000 11/06/01 (4000) 11/06/01- 27000 sampling event 11126101b,c 0 2.93 07/17/02- 8800 08/07/02 (190) 08/07/02- 8400 08/21/02 (180) 09/11/02- 9600 09/25/02 (400) 11/07/02- 11000 11/26/02 (2000) 10/01/03- 10000 10115103b, (620) 11/05/03- 14000 11/24/03c (4800) 07/08/0407/29/04b 8300 (390) 08/19/04- 10000 09/09/04bC (1700) 10/19/04- 21000 11/09/04c (4700) DO ORP NH4-N NO3-N (mg/L) (mV) (mgIL) (mg/L) 0.12 7.2 140 0.054 2.1 (0.15) (0.022) (0.44) (24) (0.097) (0.24) 20 7.4 0.11 8.6 210 0.12 2.2 (0.19) (0.17) (0.022) (0.43) (23) (0.030) (0.19) 13 7.4 0.096 11 200 0.14 1.4 (0.40) (0.13) (0.034) (0.15) (25) (0.046) (0.24) 10 7.4 0.12 12 210 0.14 1.7 (0.14) (0.066) (0.027) (0.41) (14) (0.024) (0.18) 22 7.4 0.17 8.7 160 0.25 2.1 (0.37) (0.14) (0.19) (0.63) (22) (0.14) (0.66) 22 7.3 0.19 8.3 150 0.25 2.0 (0.35) (0.17) (0.25) (0.63) (29) (0.18) (0.70) 19 7.4 0.098 9.3 160 0.21 1.6 (0.76) (0.079) (0.015) (0.44) (28) (0.021) (0.49) 9.5 7.2 0.078 12 160 0.18 1,3 (0.46) (0.11) (0.016) (0.17) (12) (0.017) (0.40) 18 7.2 0.10 10 280 0.24 0.62 (0.22) (0.18) (0.017) (0.86) (23) (0.034) (0.094) 9.0 NA NA NA NA NA NA 23 7.2 0.097 9.5 160 0.032 0.75 (0.85) (0.10) (0.011) (1.2) (28) (0.017) (0.18) 22 7.2 0.10 8.5 170 0.16 0.57 (0.26) (0.051) (0.018) (1.2) (28) (0.028) (0.12) 9.6 6.4a 0.078 12 310 0.17 0.16 (1.0) (0.13) (0.018) (0.49) (127) (0.045) (0.10) 3.86 (19000) 0.04 0 0.41 1.88 2.32 2.71 specific pH conductivity (mS/cm) (1.0) 0.04 2.35 2.00 values are average of 11 sampling sites; numbers in parenthesis are 1 SD; NA - data not available a lower pH was measured at River Mile 3.5 East (pH = 4.2) and RM Mile 7 West (pH = 5.1). There was no overt failure of the probe. These two measurements appeared to be an outlier and we did not include to the average; b sewage overflow occurred before or during sample deployment; C rainstorm during sample deployment 35 Table 2.2 Summary results of total organic carbon (TOC), dissolved organic carbon (DOC), total suspended solids (TSS), and total dissolved solids (TDS) at the Willamette River, Oregon during SPMD deployments. sampling date 11/27/01 11/26/02 10/0 1/03 11/05/03 07/08/04 07/29/04 08/19/04 09/09/04 TOC (mg/L) 2.4 (0.47) 2.2 (0.81) DOC (mg/L) TSS (mg/L) TDS (mg/L) NA NA NA NA NA NA NA 4.2 (1.2) 6.8 (1.3) 7.8 (2.6) 7.8 (6.5) 6.8 (3.4) 120 (24) 66 (9.3) 94 (30) 86 (9.0) 74 (9,7) 2.1 (0.83) 1.4 1.3 (0.0058) (0.044) 1.6 1.6 (0.058) (0.050) 1.7 1.8 (0.21) (0.27) 1.9 1.9 (0.21) (0.17) 1.7 1.8 (0.073) (0.093) 1.8 1.7 (0.076) (0.060) - Values are averages of 10 sampling sites (River Mile 1 - 18) in 2001, and 4 sampling sites (River Mile 1, River Mile 7 West, River Mile 7 East, and River Mile 18) in 2002, 2003, and 2004. Spatial differences between sampling sites were not statistically significant, therefore the data were averaged. - Numbers in parenthesis are 1 SD - NA; data not available The river characteristics and water chemistry during passive sampling device deployments are summarized in Table 2.1. In situ river physico-chemical parameters were collected at the river bottom, at 10 ft. from the river bottom where the samplers were located, and at the river surface. At each sampling site, water chemistry parameters of the three vertical levels were not statistically different within a sampling event. Also within the study area (18 miles), there was no statistical difference between the sampling sites. The physicochemical properties measured at 10 ft. from the river bottom at the eleven sampling sites were therefore averaged and reported in Table 2.1. The difference in water temperature in summer and fall was approximately 10 °C. Dissolved oxygen concentrations ranged from 7.2 mg/L in summer to 12 mg/L in fall. The river pH was neutral. The oxidation-reduction potential tended to decrease in periods of low flow in summer (p = 0.054, t-test). There was no seasonal pattern observed for the river specific conductivity, NH4tN, and NO3 -N 36 concentrations. In addition, the concentrations of TOC, DOC, TSS, and TDS did not change seasonally (Table 2.2) which suggested that a major transport of contaminants in the lower Willamette River was in the aqueous phase. The study of aqueous phase bioavailable contaminant distribution in the present study is therefore relevant to temporal river contaminant characterization. Distributions of bioavailable DDT concentrations and potential sources. Bioavailable DDT concentrations (sum of p, p'-DDT, p, p'-DDD, and p, p'-DDE; below detection limits were treated as zero) from the eleven sampling sites over the four-year study period varied from 27 pg/L to 1500 pg/L with an average concentration of 190 pgIL. The highest concentrations were always observed at River Mile 7 West (350-1,500 pgIL). The concentrations of DDTs in the Willamette River at River Mile 7 West were generally higher than dissolved DDT concentrations of several other large rivers in the US including the San Joaquin River, California (< 1,000 pgIL) (33), the Missouri River, Missouri (27-270 pgIL) (34), and the Susquehanna River, Maryland (280 pg/L) (10). However they were comparable to or much lower than the concentrations in several rivers in other parts of the world such as the Humber catchments, UK; 220 - 1,800 pgIL(35), the Minjiang River, China; 40,000230,000 pg/L (36) and the Kafue River, Zambia; 7,000-35,000 pg/L (37). It is important to note that only the studies at the Missouri River and the Kafue River used passive sampling devices to study the dissolved concentrations while the other studies used conventional filtered water methodology. The two approaches are not directly comparable since, unlike the passive sampling technique, the dissolved concentrations in filtered waters would include DDTs associated with dissolved organic matter. Therefore the non-passive sampling device technique may overestimate the truly dissolved bioavailable concentrations. However, in general, the Willamette River at Portland Harbor, particularly at River Mile 7 West is more contaminated with DDTs than many studied compromised rivers in the US, but it is generally less than compromised rives worldwide. Figure 2.2 shows the average concentrations of DDTs estimated from DDT concentrations in the SPMD. Spatial distribution of DDTs was observed both in summer and fall conditions. Bioavailable concentrations of DDTs and each individual p, p'-homolog 37 at River Mile 7 West were significantly higher than any other sampling sites in both summer and fall (p <0.001; ANOVA F-test and Tukey's HSD). The concentrations of bioavailable DDTs downstream of River Mile 7 west were apparently diluted from the concentrations measured at River Mile 7 West. The average bioavailable DDT concentrations at the upstream sampling sites including River Mile 7 East were approximately 10-fold less than those at River Mile 7 West (Figure 2.2). DDT production and use in the US has been banned for three decades. Surface water contamination would remain only if DDTs are continuously introduced through atmospheric deposition, stream runoff, and non point source inputs of surrounding soils and sediment from both in and adjacent to the water body. A contribution of DDTs from atmospheric deposition is not likely to account for a particular site-specific contamination within the 18- mile stretch. The 10-fold increases of bioavailable DDTs in surface water at River Mile 7 West, as demonstrated in Figure 2.2, indicated the upstream sources were not as significant as the local source at River Mile 7 West. Our findings (Figure 2.2) combined with lack of organochiorine insecticides, including p,p'-DDE detected in the filtered water samples from the Willamette River tributaries by Anderson et al. (38) suggest the tributary influences on the bioavailable DDT distributions in the Lower Willamette River are probably small. Data from Figure 2.2 further reveals that the ability of stream flow to transport bioavailable DDT and its metabolites during high flow periods in fall is insignificant. These findings indicate that sources upstream of the harbor are negligible for bioavailable DDTs. 38 -J -J 0) 1400 0. 0 0 0. DOTs 0 1200 0 0 1000 a) C) 800 - 0 0 600 - a) 800 0 C 0 600 0 400 Ca 200 <a > 200 > <a 0 <a 0 .0 1200 1000 400 .0 1400 0 0 :3 0. - 300 300 p,p- DOT 250 C 200 C C 150 - 200 150 - 0 0 o 100 - 100 - .0 <a p,p- DDT 250 a) 0 0 ,ck .6; :3 50 - > <a 50- 00 0 > <a 0 .6 0. çi;5 300 p,p- DDE 250 C 0. 200 C 0 a) 0 0 150 - 0 0 100- .0 50> 00 (a .0 +± + 50 <a rir1r > 00 <a .6 p,p- DDD 1000 C 0 5 \1 \'Ô p,p- ODD 800 600 - C-) 400 C) 400 :3 .9 200 0 \ C) .0 .0 -6; 800 0 600 C 0 0 > <a 0 \?; 1 a) C) 150- C) 100 0. C 200 - .0 o,1206' 1000 p,p- DDE 250 %- ", > <a 0 .0 0 ee\1 Figure 2.2 Average concentrations of bioavailable DDTs in surface water at the lower Willamette River at Portland Harbor, Oregon, using passive sampling device (n=186). Passive sampling devices were deployed for 7-21 days during the periods of low flow in summer, and high flow in fall of 2001-2004. 39 The bioavailable DDT concentrations at River Mile 7 West increased significantly during low-flow in summer sampling events, indicating a source of DDT in this area may be important. A former DDT manufacturing and handling facilities were approximately 1 mile upstream of the sampling site at River Mile 7 West. The spatial distribution of bioavailable DDT concentrations coincided with the DDT distributions found in sediment (15). Weston et al. (15) reported surface sediment samples collected within 1 mile downstream of the former DDT facilities contained DDT concentrations approximately 4- to 10-fold higher than in the samples upstream or downstream. Historically discharged DDTs in sediment could be recycling into the water column via the contaminated sediments (39). Contaminants that are dissolved or associated with colloidal particles can exchange across sediment-water interface by diffusive or advective processes (40, 41). Subsequently, the sediments as a depositional reservoir for historically discharged contaminants could have become an important source of contamination and continued to recycle deposited contaminants to the overlying water column (42). The presence of bioavailable p, p'-DDT and its metabolites presumably resulted from historically DDT-deposited sediments. The findings suggest that historically contaminated sediments rather than external non-point sources have become the dominant source of DDT contamination in Portland Harbor. Dominance of bioavailable p, p'-DDD in surface water. The dominance of bioavailable p, p'-DDD over bioavailable p, p'-DDE was observed. Figure 2.2 reveals p, p'DDD is the most abundant bioavailable DDT derivatives followed by p, p'-DDE, and p, p'DDT. The average percentage contributions of p, p'-DDD, p, p'-DDE, and p, p'-DDT to DDTs in the samples from eleven sampling sites in four years were 53 ± 14%, 35 ± 11%, and 13 ± 6%, respectively. In general, the principal insecticidal ingredients of technical DDT contained p,p'-DDT 72%, o, p'-DDT 20%, p,p'-DDD 3%, o, o'-DDT 0.5% and other 4.5% (43). Both p, p'-DDD and p, p'-DDE existed as by-products in technical DDT and have been formed by environmental degradation of DDT. The high contribution of p, p' -DDD and p, p' DDE in the bioavailable fraction suggested a large proportion of p, p'-DDT has been transformed to p, p'-DDD and p, p'-DDE. It is noteworthy that p, p'-DDD was also marketed as an insecticide in its own right, as well as, being a reductive metabolite of p, p'-DDT (43). In addition to being a reductive metabolite of p, p' -DDT, a large proportion of p, p' -DDD 40 could therefore indicate a direct input of p, p'-DDD in this area. The results indicated p. p'DDD was the most bioavailable p. p'-homolog in surface waters of the lower Willamette River The large bioavailable concentrations of DDTs coincided with large DDD/ DDE ratios (Figure 2.3). The ratio of DDDIDDE was highly variable between sites and sampling periods. The DDDIDDE ratio at River Mile 7 West was highly variable, 6.6 ± 2.1 in summer, and 2.8 ± 1.3 in fall. The large proportion of bioavailable p, p'-DDD and the large ratio of bioavailable DDD/DDE in surface waters could be governed by water solubility and the partition coefficients (log K0 and log K). Water solubility of p, p'-DDT, p. p'-DDD and p,p'-DDE are 0.0055 mg/L, 0.02 mg/L, and 0.1 mg/L at 20-25 °C (44). However, the bioavailable p. p'-DDT, p. p'-DDD, and p. p'-DDE concentrations in this study were measured at the level much below the water solubility and therefore water solubility is unlikely to be the cause of the observation. Log K; 5.7 for p,p'-DDT, 6.1 for p. p'-DDD. 6.0 for p. p'-DDE are similar, as well the sorption coefficients (log K); 6.3 for p. p'-DDT, 5.0 for p. p'-DDD, 4.7 for p. p'-DDE at 20-25 °C (32, 44) are not likely to account for the observation seen. It is unlikely that the large proportion of bioavailable p. p'-DDD and large ratios of bioavailable DDD/DDE are the results of differences in solubility or partitioning, suggesting the large bioavailable p. p'-DDD concentrations may be influenced by other factors. 10 0 6- aa aa 4a 2- 0 3? o 0 3 0 o VT V 0 summer faU 0 (5 w o v 0 0 8- 0 0 ill 0 0 V 0 0 0 0 0 V Figure 2.3 Distributions of bioavailable p. p'-DDD: p. p'-DDE ratios in surface waters at the Willamette River, Oregon during the periods of low flow in summer, and high flow in fall from 2001 to 2004 (n=186). 41 The large ratio of DDD/DDE was previously observed in the Portland Harbor sediments from River Mile 7-8 West (15). Zeng and Tran (42) showed evidence for a strong tendency for organochiorines including DDTs, to move from sediment to overlying water. The similarity of DDT profiles in the sediment (15) and the water bioavailable fraction supports the possibility of the movement of bioavailable DDT and its metabolites to the water column from historically contaminated sediments. The large ratio of DDD/DDE could also indicate the potential for reductive dechlorination of DDTs to DDDs in sediments under flooded, anaerobic conditions (45, 46). Under anaerobic conditions in sediments, DDT is mainly metabolized to DDD by reductive dechlorination either by microbial degradation or by chemical reaction while under aerobic condition, DDT is metabolized to DDE by dehydrochlorination (47-49). It is possible that the lower Willamette River at Portland Harbor which is a deep and slow moving river (15) has a potential condition for anaerobic reductive dechlorination of DDT to DDD rather than to DDE. In general, p, p'-DDE tends to resist further degradation as compared to p, p'-DDD (48, 49). However, Quensen et al. (50) found that in anaerobic marine sediments, DDE was dechlorinated to DDMU (1-chloro-2,2-bis-(4'chlorophenyl)-ethylene) three orders of magnitude faster than the transformation of DDD to DDMU. In addition to direct input of DDD, anaerobic reductive dechlorination of DDT to DDD and possible persistence to further degradation of DDD under anaerobic condition in sediments may all be responsible for the enrichment of bioavailable p,p' -DDD in Portland Harbor. However, further investigations of dechlorination conditions in the harbor sediment are warranted. Evidence of seasonality in bioavailable DDT concentrations. Seasonal variations of bioavailable DDTs at the lower Willamette River at Portland Harbor were observed. Seasonal variation of bioavailable DDTs is site-specific. Seasonal variation of bioavailable DDT concentrations was only statistically different at River Mile 7 West (p = 0.002, t-test) (Figure 2.2). A 2-fold decrease of bioavailable DDTs during periods of high flow in fall was measured at River Mile 7 West. The decrease in fall is due to a decrease of bioavailable p, p'- DDD. In contrast, bioavailable p, p'-DDT and p, p'-DDE did not decrease in fall. By contrast to River Mile 7 West, bioavailable DDT concentrations at the other sites remained unchanged between summer and fall. 42 30 water temperature 25 - :: 2015 10 - = 5000k 30000 Co .2 a) > river flow 20000 - I 10000 - e =1=1 I I precipitation 4 3 2 0 0) E 16 14 12 10 0 a 8 6 dissolved oxygen 0 § 0 0 4 50 p,p-DDT 400 300 200 100 iii 1 4OB 1050 700 350 0 500 400 = 300 200 100 - p 'p -D D D lk;I p ,p-D D E 0 10 p,p-DDD: p,p -DDE ratio 8= 642- w 0 O0 c' c°' e0 2.c0 00200 o ,Zc30 ec'° ee0 c'° e 0 sam pling event Figure 2.4 Estimated concentrations of bioavailable p, p'-DDT and its metabolites in surface water at River Mile 7 West of the lower Willarnette River at Portland Harbor, Oregon in relation to average water temperature, average river flow, total precipitation, total organic carbon (TOC, black circle), and dissolved organic carbon (DOC, white circle) concentrations. 43 At River Mile 7 West, bioavailable DDT concentrations were significantly lower in fall (p = 0.002, t-test), excluding October 2004 (Figure 2.4). Bioavailable p, p'-DDT, p, p'DDD, and p, p'-DDE concentrations in October 2004 were unusually high compared to the concentrations measured in fall 2001 to 2003 and were treated as an outlier in this case, In general, a significant decrease of bioavailable p, p'-DDD (p = 0.001, t-test) in fall appeared to be a major driver of seasonality in bioavailable DDT concentrations. l3ioavailable p, p'DDT and p, p'-DDE were statistically unchanged between seasons (p = 0.58 for p, p'-DDT andp = 0.50 for p, p'-DDE, t-test). The bioavailable p, p'-DDD concentration was positively correlated with average water temperature (r2 = 0.62, p = 0.003), and negatively correlated with average river flow (r2 = 0.39, p = 0.03) and total precipitation (r2 = 0.71, p <0.001) (Figure 2.4). As dissolve oxygen concentration is a function of water temperature, bioavailable p, p'-DDD was also negatively correlated with dissolved oxygen concentration (r2 = 0.49, p = 0.0 16). Bioavailable p, p'-DDD concentration was not statistically correlated to total organic carbon (TOC) and dissolved organic carbon (DOC) concentrations (Figure 2.4). The results suggest seasonal change in bioavailable p, p'-DDD concentrations is potentially related to changes in temperature, river flow, andlor precipitation. The average amount or load of bioavailable DDTs transported in surface water at the lower Willamette River (River Mile 1-18) during the 4- year study in summer ranged from 1.1 g/d to 22 g/d for bioavailable DDTs; 0.11 g/d to 2.1 g/d for bioavailable p, p'-DDT; 0.61 gld to 18 g/d for bioavailable p,p'-DDD; and 0.36 gld to 2.7 g/d for bioavailable p,p'- DDE. The average loads during fall were 2.6 g/d to 27 g/d for bioavailable DDTs; 0.26 g/d to 6.9 g/d for bioavailable p, p'-DDT; 0.86 g/d to 15 gld for bioavailable p, p'-DDD; and 1.3 g/d to 6 .0 g/d for bioavailable p, p'-DDE. The highest loads of bioavailable DDTs always observed at River Mile 7 West during summer and fall. In general, the loads of bioavailable p, p'-DDT and its metabolites were not significantly different between seasons (p > 0.05, ttest). Daily average river flow and water temperature were used to evaluate the seasonality of bioavailable p, p'-DDD concentrations and ratios of bioavailable DDD/DDE (Figure 2.4). An increase of bioavailable p, p'-DDD concentrations occurred during the low flow in summer with a maximum observed during extended dry periods. Simply, one might be 44 tempted to think dilution could account for the decrease in bioavailable p,p' -DDD concentrations during fall sampling events when the river flow is large. However, if dilution was the cause, dilution should decrease all analytes similarly; this was not what we found. Both bioavailable p, p'-DDT and p, p'-DDE were found to have the same or higher concentrations in fall as compared to summer. Also, the DDD/DDE ratio should not change if dilution is a major cause. It appeared that physical-chemical processes governed by water temperature and river flow variations alone could not explain the observed seasonal variation in bioavailable p, p'-DDD. We hypothesize that the seasonal distribution of bioavailable p,p'DDD is governed by a combination of physical and biological factors that vary seasonally as a function of temperature, flow velocities, and aquatic organism activities. The occurrence of elevated bioavailable p,p'-DDD concentrations and bioavailable DDD/DDE ratios in mid July to mid September coincided with elevated temperature. Increase in water temperature could decrease the organic matter-water partition coefficient (Kom) of contaminants (51). Schwarzenbach et al. (51) estimated increasing temperature by 10 °C could result in an estimated 30% decreased absorption coefficient for pyrene. The effect of increasing temperature by approximately 10 °C in summer could result in an increased dissolved fraction from the sediment-water exchange for DDTs. A 30% increase in bioavailable DDTs release would result in elevated concentrations of DDTs in surface water. However, it appeared that increase of DDTs release from the contaminated sediment due to decreasing temperature-dependent absorption coefficients alone might not readily explain the significant increase in DDD/DDE ratios observed in summer. This gap suggests another influencing factor(s) that could contribute to the elevated concentrations of bioavailable p. p'-DDD in summer. Increased water temperature would also be expected to increase growth and activity of benthic organism, microorganisms, eutrophication, and bitoturbation. Bioturbation by burrowing organisms at the surface sediment can reintroduce sediment-borne chemicals to the water column (52). Some aquatic plants can uptake and transform p, p'-DDT primarily to p, p'-DDD (53). Increasing aquatic plant growth could lead to oxygen depletion as microorganisms break down the dead tissues. The top layer of the sediment may be aerobic or anaerobic depending on the condition of the water column above; however the sediments 45 quickly become anaerobic with increasing sediment depth (52). There was also declining redox condition (p = 0.054, t-test) as water temperature and dissolved oxygen concentrations changed in summer (Table 2.1), a contributing factor to enhance anaerobic condition in the sediments. Anaerobic reductive condition is the dominant condition for reductive dechlorination of p, p'-DDT top, p'-DDD by both microbial degradation and chemical reaction (47-49). Slow moving surface waters during summer would enhance anaerobic reductive degradation to p, p'-DDD as Kale et al.(45) found p, p'-DDD was a major metabolite of p, p'-DDT detected in sediment and overlying water under flooded condition. Evaluated together, increasing formation and release of p, p'-DDD as result of increased temperature, decreased dissolved oxygen concentration, and the condition favoring reductive conditions in sediment during low flow in summer would contribute to an increase of bioavailable p,p'-DDD and high ratio of DDD/DDE in surface water. As river flow and precipitation increased in early fall, the formation and release of p, p'-DDD could be affected by dilution process as suggested by relatively no change in DDDIDDE ratio. Decreasing ratio of DDD/DDE in fall suggested seasonal influence on p, p'-DDD sources. As water temperature decreased and river flow increased in fall, the condition suitable for reductive dechlorination may have been altered resulting in decreased bioavailable p, p'-DDD concentrations and change in bioavailable DDDIDDE ratios. The above discussion contains speculation of the processes that highlights the lack of understanding on how seasonal river conditions interact to yield the observed seasonal variation in bioavailable p, p'-DDD and its relationship to bioavailable p, p'-DDE. Further research is needed. Understanding the interaction of seasonal physico-chemical and biological factors will afford confidence in using this knowledge to predict the long-term seasonal effects of DDTs on aquatic community and increased confidence in extrapolating information to other contaminated sites. Potential risk of seasonal bioavailable DDTs on aquatic organisms. The present study measured the bioavailable concentrations of DDTs in surface water which allowed a direct assessment of the water quality in the Portland Harbor. Aquatic organisms can take up large amount of DDTs over time from surrounding water where DDTs are present at very 46 low concentrations (54). It has been addressed that DDTs taken into aquatic organisms has come from sunounding water while a large proportion of DDTs in their body has come from the food (54). To assess the potential impacts of seasonal distribution of DDTs on aquatic organisms at the lower Willamette River, water concentration estimates were compared to the national recommended water quality criteria (22) and the Oregon water quality criteria (23) (Figure 2.6). Exceedances of the fresh water aquatic life criteria for DDTs (1,000 pgJL for both national and Oregon criteria) were seasonally influenced and limited to the samples from RM 7 West. The bioavailable concentrations of bioavailable DDTs always exceeded the national and the Oregon criteria during low flow in summer. During sunmier, the aquatic organisms at RM 7 West could have been exposed to a greater amount ofDDTs, particularly p, p'-DDD, due not only to seasonally increased exposure concentrations but also to increased uptake rate. As aquatic organisms are ectothemic, the rate of metabolism undergoes an approximately 2-fold increase with every 10 °C rise in water temperature (55). Cairns et al (55) suggested increased temperature could change respiratory rate and membrane permeability and thereby leaded to more rapid uptake and hastened the accumulation of a toxic dose. The effect of increasing temperature approximately by 10 °C in summer could result in increased DDT bioconcentration from surrounding water. Water concentrations determined in this study are the bioavailable fraction which has been known as the uptaken form by aquatic organisms. Therefore, exceedance of DDT fresh water aquatic life criteria, as well as, the potential to increase their ability to bioconcentrate DDTs from surrounding water during warm weather can pose a potential risk for DDT chronic toxicity to aquatic communities at River Mile 7 West. 47 1600 0 1400 0 S 1200 o 1000 O National hesh Water aquatic life criteria and Oregon criteria 0 A a) 60003 0 .0 400 - 200- t o. 350 I0 ç .0 8 0 in in 150 ii too 0 50 > 0 0 116*011 0 0 800 a a 8 600 0 400 _iO 0 0 200 5hea0hcr5ena pp-DDE 350 0 at (it Oregon human 0 - 400 0 1200 1000 a National human health water quality criteria 450 p,p-DDD 1400 0 300 250 0. 200 at 0 1600 - p,p-DDT A 400 a 800 - 450 DDTs Netional human health water quality csteria .0 .5 .0 300 250 Netionel human health water qualrty criteria A £ 200 S t50 0 100 I 0 0 summer A fall Figure 2.6 Spatial and seasonal distribution of bioavailable p, p'-DDT and its metabolite water concentration estimates compared to the national recommendation water criteria (22) and Oregon criteria (23) to protect aquatic organism from chronic exposure to DDTs and to protect local fish consumers who consume fish and shellfish from the lower Willamette River at Portland Harbor, Oregon. Bioavailable concentrations of p. p'-DDT, p, p'-DDD, and p, p'-DDE were also compared to the national and Oregon human health water quality criteria to estimate life-time cancer risk for local fish consumers based on a 106 increased lifetime cancer risk (Figure 2.6). Exceedances of the human health water quality criteria for p. p'-DDT and its metabolites were mostly observed at RM 7 West. Although no samples exceeded the national criteria for p. p'-DDT (220 pgIL), all samples from RM 7 West exceeded the Oregon criteria (24 pgIL). Exceedance of the national human health water quality criteria for p, p'-DDD (310 pgIL) was seasonally affected. Bioavailable concentrations of p. p'-DDD tended to exceed the criteria during low flow in summer sampling events. Only a few samples exceeded the criteria for p. p'-DDE (220 pg/L). Although, the human health water quality criteria is not the direct approach for human health risk assessment for fish consumers but it is useful as a screening approach for indicating potential areas of concern. According to the results in our 48 study, the potential adverse effects ofDDTs for local fish consumers were realized locally at River Mile 7 West. This finding agreed with the bioaccumulation profiles of DDTs in fish and human health risk estimates for DDTs (2 x 106 to 8 x 10-s) from consuming fish in this area in our previous study (17, 18). The excellent agreement between the present study and our previous fish study confirmed passive sampling devices can be used as a surrogate to study bioavailable distribution of organic contaminants in contaminated water. Distribution of bioavailable potychiorinated biphenyls (PCBs). Iii contrast to the generally point source nature of DDT contamination, there were several former PCB using facilities densely located on both sides of the river from RM 2-10.5 (56). These facilities have been identified as potential sources of PCB contamination in the harbor. Elevated concentrations of PCBs were detected in sediments and subsurface sediments from RM 3.5 to 9.5 (15). Dissolved PCBs or colloidal particle-associated PCBs in the sediment can exchange across sediment-water interface by diffusive or advective processes, subsequently, recycling historically deposited PCBs to the water column (40-42). In addition, the current major sources of PCBs to urban rivers include direct urban runoff, urban community wastewater discharge, sewage treatment works, and combined sewer flow discharges (5759). Considering the influence of multiple PCB local sources and upland land uses, the sampling sites were then grouped into three locations; RM 1-8 (industrial area including most of Portland Harbor superfund site), RM 12-13 (urban area) and RM 15-18 (residential area). The spatial pattern of bioavailable PCB concentrations in water coincided with the historically spatial PCB sediment distribution. A spatial pattern of bioavailable PCB concentrations was observed in summer and fall (Figure 2.7). Bioavailable concentrations of PCBs ranged from below detection limits (0.69-54 pg/L depending on congeners) to 410 pg/L at the eleven sampling sites from River Mile 1-18 during the four-year study with an average concentration of 54 ± 55 pgIL. In general, higher bioavailable PCB concentrations occurred in the industrial and urban areas from River Mile 1-13. Elevated concentrations of bioavailable PCBs usually reached a maximum at RM 3.5 East, the lower superfund site. Average bioavailable concentrations of PCBs during low flow in summer ranged from 47 74 pg/L at the industrial area, from 26-33 pg/L at the urban area and from 12-15 pg/L at the residential area. Average bioavailable concentrations of PCBs during high flow in fall 49 ranged from 37-170 pgfL at the industrial area, from 36-69 pgIL at the urban area and from 23-42 pg/L at the residential area. i 320 -S 88 a. 150- N 100- 500 0 i-i-I lEi - Summer Fall s Figure 2.7 Averaged concentration estimates of bioavailable PCB (pgIL) in water at the Willamette River, Oregon according to river season (low flow, low precipitation in summer and high flow, high precipitation in fall) in 2001 to 2004. The error bars denote 1 SD. The bioavailable concentrations of PCBs in the lower Willamette River were generally comparable with or lower than the dissolved concentrations determined in other large water bodies in US including San Diego Bay (11 - 330 pg/L based on 27 PCB congeners) (60), Lake Superior (90 ±10 pg/L based on 25 PCB congeners) (61), Lake Michigan (340 -1700 pg/L based on 85 PCB congeners) (62), the Susquehanna River, Maryland (500 -5300 pg/L based on 85 individual congeners) (10), and much lower than those in some river systems around the world such as Minjiang River, China (200,000 2,500,000 pg/L based on 21 PCB congeners) (36). It is noted that comparison between PCR contamination at the lower Willamette River and the other rivers did not account for the variations from sampling technique used, the techniques used to extract the operationallydefined dissolved phase and numbers of congeners analyzed. Other techniques probably overestimate the bioavailable concentrations. 50 Seasonal changes in bioavailable PCBs distributions. Bioavailable concentrations of PCBs showed a discernible seasonal pattern at almost every sampling site (Figure 2.7). A two-fold increase in bioavailable PCB concentrations was observed during the high flow in fall, with the residential site (RM 15-18) statistically significant (p = 0.003, t-test). The average daily amounts or loads of bioavailable PCBs transported in surface water at the lower Willamette River also showed significant seasonal differences. Average of bioavailable PCB daily loads in surface waters were significantly higher in fall; industrial area (p 0.002, t-test), urban area (p = 0.043, t-test), and residential area (p = 0,001, t-test). The average of bioavailable PCB daily loads at the industrial area (River Mile 1-8) during high flow in fall was 3.4 g/d, compared to 1.2 g/d during low flow in summer. At the urban area (River Mile 12-13) and residential area (River Mile 15-18), the average of bioavailable PCB daily loads during high flow in fall was 2.56 g/d and 1.46 g/d, compared to 0.62 g/d and 0.28 g/d during low flow in summer. In spite of the dilution effect, bioavailable water PCB concentrations were larger during high flow in fall than during low flow in summer. Bioavailable PCB composition in surface water. The average percent congener distributions of bioavailable PCBs are shown in Figure 2.8. PCB 153 and PCB 138 (hexa-CB homolog) were the most frequently detected bioavailable congeners with 85% and 73% frequency, respectively. PCB 49 (tetra-CB) was the most abundant bioavailable congeners followed by PCB 52 (tetra-CB) and PCB 153 (hexa-CB) as the second most abundant bioavailable congeners detected. PCB 166 and two of the co-planar, dioxin-like congeners, PCB 77 and PCB 126 were below the detection limits in all samples. 51 0 ° 60 60 50 50 40 40 30 0 20 O 10 0 N-)('4QrC) N-N 0 a U) )0(DO)O0C) N. C) 00 10 0 N- 0 C)CJOON- N- 60 60 50 50 40 40 30 0 o C C U) 10 U) 0 a 20 CC o 20 0 30 U) 0 0 C)N0)00N-U)0C)0 N-' CCC 0)00CC N. 0) N-N- 00 OOCCN. C) 30 20 10 a0)0 N.0 00N.N-C) N-N00 CCC)OOCCN-C) 60 50 40 30 o0 20 0U) 10 a 0 U) N0CCO(Da)OOCC N-C) congener no. N-0O)C'JO0 N- N-C)CC N-N- 00 00CC N-C) congener no. Figure 2.8 Average percent contributions of PCB congeners in surface water at the lower Willamette River during the periods of low flow in summer and high flow in fall in 2001 to 2004. The error bars denoted 1 SD. Although bioavailable PCB concentrations and loads were found to have strong spatial and seasonal variation, the PCB congener profiles did not. The congener profiles for River Mile 1-8 and River Mile 12-13 were fairly uniform and no discernable seasonal patterns were observed, Figure 2.8. The similarity of PCB congener profiles for the industrial area at the superfund site (River Mile 1-8) and for the urban area upstream (River Mile 1213) suggests the local sources of PCBs at the superfund site are important enough to significantly increase the concentrations of bioavailable PCBs in surface water but not to substantially change composition relative to the surface water inputs from upstream. 52 The average percent congener distributions for River Mile 15-18 relative to the average of the other sampling sites showed the bioavailable PCBs to be comprised of a slightly different composition, with the distribution maximum shifted toward the hexa homolog (PCB 138 and PCB 153) as compared to the other sampling sites, which favored the tetra homologs (PCB 44, PCB 49 and PCB 52), Figure 2.8 and 2.9. However, the average percent congener and homolog distributions at River Mile 15-18 became similar to those at the other sampling sites in the periods of high flow in fall. g 10 0) C) River Mile 1-8: summer 0.8- River Mile 1-8: tall 0.4- 04 o 02E a 02 E g 00 C) 0 0.0 -CS 1.0 River Mile 12-13: summer 0) , 08 C) N 0 o E 0 -z 06 N 0.4 - 0) o 0.2 E 0 10 River Mile 15-18: summer 0) o C) 0.8 a 02 I 0 N C) 0 0 E GSva GCB G5C5 River Mile 15-18: laO 0) 0 T E 00 0 10 C) 06 04 0.4 0 0.2- 0 080.6 0.4 - 0200 ,T I'] Figure 2.9 Average ratios of PCB homolog to bioavailable PCBs (sum of 25 individual PCB congeners) in surface water at the lower Willamette River during the periods of low flow in summer and high flow in fall in 2001 to 2004. The error bars denoted 1 SD. See detail of PCB homologs in text. The present study suggests the sources of bioavailable PCBs in surface waters at the lower Willamette were presumably dependent on seasons. Storm flow was especially important. Both concentrations and daily loads of bioavailable PCBs increased during high 53 precipitation and high river flow, especially during episodic rainstorms in November of 2001 and October of 2004. The importance of high river flow enhancing PCB transport in surface waters has been previously described (10). Foster et al. (10) demonstrated PCB sorption coefficients (Kds) were decreased during storm flows and subsequently enhanced the dissolved, or the "bioavailable" PCB concentrations in the surface waters. They indicated the lower Kjs of PCBs during storm flows may be attributed to an increase in DOC, a change in particle organic matter content, or shifts in PCB congener profiles to a greater extent of the less chlorinated congeners. In the present study, DOC and TOC concentrations at the lower Willamette River did not change seasonally over the 4 year-study period (Table 2). DOC and TOC concentrations were not correlated with the bioavailable PCB concentrations. There was no evidence of seasonal or river flow-related change in PCB congener or PCB homolog profiles observed at River Mile 1-8 where an average 50% increase in bioavailable PCB concentrations was observed. In contrast, a shift in PCB congener and homolog compositions toward the less chlorinated congeners was observed at River Mile 12-13 and River Mile 1518. High river velocities during storms can promote the transport of a relatively larger fraction of course particles in particulate phase which have lower organic carbon contents (i.e., changing organic matter content) and reduced PCB sorption(63). The results suggests storm events (i.e., high river flow, high precipitation) may have disturbed the interaction between PCBs and their associated organic matters resulting in remobilization of bioavailable fraction from sediments or particulate matters. Additional inputs during high flow in fall such as precipitation and stoi in water runoff may have also contributed to elevated bioavailable PCB concentrations and PCB composition changes during high flow in falls. Atmospheric transport including precipitation is an important pathway for the transfer of PCBs to surface water as indicated in several studies (12, 64, 65). Bremle and Larsson (12) reported precipitation was a dominant source of PCBs in a Swedish river during high flow. The PCB congener and homolog profile in surface water samples from the lower Willamette River were similar to that observed in rain samples in other studies, which were dominated by tetra-chiorinated congeners, including PCB 52 (64, 65). The similarity of PCB homolog patterns in our study and those in rain samples by others may suggest precipitation inputs as an additional potential source of PCBs into the river system during high precipitation and further investigation is needed. 54 Urban storm waters and urban community wastewater discharges are another source of PCB contamination in the surface waters (57 59). Rossi et al (57) reported urban storm water was a major contribution to PCBs in Swiss urban water systems. In the New York/New Jersey Harbor Estuary, USA, PCB concentrations in the influent were elevated during storrns(59). Approximately, 3% of the annual PCB contribution from the wastewater treatment plants were diverted and bypassed the wastewater treatment plants and discharged through combined sewage overflow due to precipitation events(59). In the lower Willamette River at Portland, combined sewer overflows to the Willamette River occur frequently (i.e., 100 days or 50 events overflow in 2004), especially during periods of high precipitation which usually start in mid October(66). The increase in bioavailable PCB concentrations and loads which was coincident with high precipitation and sewer overflows (Table 2.1) may suggest a significant contribution of PCBs from urban storm waters and urban community wastewater discharges to the surface waters. The hypothesis that combined sewer overflows and precipitation could be a major source of PCBs during high precipitation and high river flow in the lower Willamette River warrants further investigation such as PCB fingerprints to fully elucidate their contribution to PCB contamination. Further studies will be necessary to estimate the impact of the internal sources (i.e., contaminated sediments) and other sources (i.e., precipitation, runoffs, wastewater discharges) of PCBs and how much they contribute to the PCB problem in the system in order to reduce the PCB load and risk to the aquatic organism community. PCBs-associated risks to aquatic community and human health. None of samples exceeded the national and the Oregon fresh water aquatic life criteria for PCBs (14000 pgIL). In contrast, exceedances of the national (64 pg/L) and the Oregon human health water quality criteria (79 pg/L) for fish consumers were often observed at the sampling sites within the superfund site (River Mile 1-8) (see Figure 2.10) but no discernable seasonal pattern was observed. Year-round exceeding the human health water quality criteria suggests consuming fish and shellfish from this area may pose adverse health effect risk to recreational or subsistence fishers based on a 106 increased lifetime cancer risk for PCBs. This suggestion is consistent with risk estimates (9 x 106 to 5 x 10) for PCBs from consuming fish in this area in our previous human health risk assessment study (17). The extent of exceedance would have been greater if more PCB congeners were included for the measurement. 55 Nevertheless, this exceedance indicates that PCB contamination remains a significant problem at the Portland Harbor superfund site. 450 A 400) 350 - 300A a. Q > 250200 150- A A 0 O A 0 A 100- £OA©*0 0iuhiiiii 50 A Oregon human health criteria National human health criteria o summer A fall Figure 2.10 Spatial and seasonal distribution of bioavailable PCB water concentration estimates (sum of 25 individual PCB congeners) compared to the national recommendation water criteria (22) and Oregon criteria (23) to protect local fish consumers who consume fish and shellfish from the lower Willamette River at Portland Harbor, Oregon. Seasonal changes in bioavailable dieldrin distribution. Bioavailable dieldrin was detected at all sampling sites. Over the 4-year study period, the ranges of bioavailable dieldrin concentrations in the mainstreams of River Mile 1-18 of the lower Willamette River were from 10 to 140 pgIL. No discernable spatial pattern was observed for dieldrin. However, river condition changes between low flow in summer and high flow in fall appeared to influence the distribution of bioavailable dieldrin in the surface water. An increase in bioavailable dieldrin concentrations was found at most sampling sites during periods of high river flow, high precipitation in fall (p 0.046, 1-test), Figure 2.11. Bioavailable dieldrin daily loads were also found to be higher in fall. The average daily loads of bioavailable dieldrin ranged from to 1.5 to 3.8 gld during high flow, compared to 0.63 to 1.3 gld during low flow. The increase in both bioavailable concentrations and loads of dieldrin were 56 coincident with high precipitation and high river flow. It is noted that the increases were higher at the sampling sites upstream, particularly at RM 18 which was downstream of Johnson Creek, an agricultural/urban creek. The seasonal pattern of bioavailable dieldrin distribution illustrates the transport behavior for non-point source contaminants which are widely dispersed and have no intentional seasonal application cycle such as with the current use pesticides. Dieldrin was widely used in the Willamette basin before it was banned and Johnson Creek has been found to be one of the most dieldrin contaminated areas in the watershed (67). Elevated concentrations of bioavailable dieldrin during high flow and storm events in fall suggests bioavailable dieldrin transport from contaminated site upstream including erosional inputs to the river during precipitation runoff from land sources or riverbed sediment (68). 0. 125 100- D -o Ca 7550- 25- > 0 -o I 5NeSaSe5aS I I'iru' c5 11 S summer J a- 150 a 125 - ioo92 I 50--5 9225> 0 -o A ii 5NBNeaS - a a a a Oregon human health criteria A - National human health cntena 6 B t:k:3 o summer A.fill Figure 2.11 Seasonal distribution of bioavailable dieldrin in surface water at the lower Willamette River at Portland Harbor, Oregon during the periods of low flow in summer and high flow in fall of 2001 -2004. The top panel shows average concentration estimates of bioavailable dieldrin while the bottom panel shows their distribution compared to the national recommendation water criteria (22) and Oregon criteria (23) to protect local fish consumers who consume fish and shellfish from this area. 57 Bioavailable dieldrin concentrations were compared to the national recommended water quality criteria (22) and the Oregon water quality criteria (23). None of samples exceeded the national (56000 pgIL) and the Oregon fresh water aquatic life criteria (1900 pgIL) for dieldrin. Exceedances of the national and the Oregon human health water quality criteria for dieldrin were frequently observed in fall, particularly at the sampling sites upstream. This finding is consistent with risk estimates (2 x 106 to 3 x l0-) for dieldriri from consuming fish in this area in our previous human health risk assessment study (17). Acknowledgment. This study was partially funded by the Society of Environmental Toxicology and Chemistry (SETAC) Early Career for Applied Ecological Research Award sponsored by the American Chemistry Council to K.A.A, the Oregon Department of Environmental Quality for the McCormick & Baxter superfund site project 2002-2003, and the Oregon Health Science University pilot project for the NIEHS/EPA Superfund Basic Research Grant. The Royal Thai government scholarship to D.S. is acknowledged. We are grateful for field assistance by R. Grove of USGS, Forest and Rangeland Ecosystem Science Center, Corvallis, OR. We would like to express appreciation for field and laboratory assistance of colleagues at Food Safety and Environmental Stewardship Program (FSES), Department of Environmental and Molecular Toxicology, Oregon State University, especially E. Johnson, G. Sower, and S. Visalli. Literature Cited Walker CH, Hopkin SP, Sibly RM, Peakall DB. 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Res. 25:1379-1389(1989). 63 CHAPTER 3 FIELD TRIAL OF TRIOLEIN-FREE LOW-DENSITY POLYETHYLENE MEMBRANE AS AN IN SITU PASSIVE WATER SAMPLER Sethajintanin, D', Johnson, E.R. 1 1,2 and Anderson, K.A 1,2 Department of Environmental and Molecular Toxicology 2 Food Safety and Environmental Stewardship Program, Oregon State University, Corvallis, Oregon 97331 64 Abstract Responding to a growing need for an inexpensive and simple time-integrative sampling device for bioavailable hydrophobic toxic contaminants in water, a triolein-free low density polyethylene membrane lay flat tubing (LFT) has been further developed. The LFT sampler is similar to semipermeable membrane device (SPMD) based on the diffusion of dissolvedlbioavailable target hydrophobic compounds through the aqueous boundary layer and polyethylene membrane and the subsequent accumulation in the membrane and a neutral lipid reservoir (i.e., triolein), mimicking uptake by living organisms. Unfortunately, the interference stemming from the triolein impurities in SPMD has sometimes impeded the analyses and some target analytes are sacrificed in the removal processes of triolein impurities. Here, we demonstrate laboratory and field verification that the LFT without triolein proved reliable and had the same benefits as SPMD, but was simpler, inexpensive and lacked interference from the triolein impurities. A total of 370 LFTs and SPMDs were deployed as a paired study at thirteen sampling sites in three different river conditions, 21day deployment, at the Portland Harbor superfund site, Oregon. LFT has shown the ability to detect polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and organochiorine (OC) pesticides as efficiently as the SPMD. Its sampling efficiency suggested the neutral lipid reservoir (triolein) present in SPMD was not generally necessary. Also, biofouling was not a problem for its field application as compared to SPMD. This simpler device tended to accumulate compounds with high log K0 faster than SPMD. LFT sampling rates were estimated and modeled for 33 target analytes, including PAHs, PCBs, and organochlorine pesticides. Approaching equilibrium appeared to be not a concern even at a superfund site. The shortest duration estimated for compounds with log K0 4.4 to maintain in linear uptake phase during LFT deployment is 24 days at water temperature 22 °C. A method for spiking permeability/performance reference compounds as an indicator for the effects of environmental conditions on the device uptake rate was achieved by using a simple spiking method in a small volume of toluene. The successful determination of field derived data illustrates the effectiveness and reliability of LFT for environmental monitoring. 65 Introduction A wide range of passive sampling devices (PSDs) has been widely used in recent decades for monitoring exposure and assessment of contaminants in water, air, and soils(111). Semipermeable membrane device (SPMD) (1), solid-phase microextraction (SPME)(2), diffusive gradients in thin-films (DGT) (3), and other sampling foiiiiats with thin polymer coatings (4, 8) are examples of passive sampling technique developed and used for environmental assessment. The generally accepted major advantage of passive sampling technique over the conventional approach is the ability to distinguish between freely dissolved and bound molecules and focuses on the availability and activity rather than the mere presence of chemicals (12). The freely dissolved fraction is environmentally relevant to chemical bioavailability, toxicity, mobility and degradation process(13, 14). Instead of measuring the total concentration in a medium, the passive sampling technique measures the concentration in a reference phase which can be brought into equilibrium with the medium(12). The availability of a chemical is measured based on the chemical potential which is logarithmically related to its fugacity and linearly related to its freely dissolved concentration in a particular medium (12). The semipermeable membrane device (SPMD) introduced by Huckins et al.(1) is a passive sampling technique that has been demonstrated to linearly sequester a variety of nonpolar and moderately polar organic contaminants (15, 16). SPMD has rapidly gained wide acceptance and has become commonplace for integrative sampling of environmental contaminants in water, air, and even soil (2, 5, 7). The device consists of low density polyethylene lay flat tubing containing a thin film of a neutral lipid, 95% pure triolein (1,2,3-tri{cis-9-octadecenoyl]glycerol) (15, 16). It has been proposed that SPMD mimics key mechanisms of bioconcentration including diffusion through biomembranes and partitioning between organism lipid and the surrounding medium (15, 16). Although SPMD provides many attributes such as a chemical bioavailability and simplicity of sample deployment and sample extraction, analysis of SPMD has sometimes been severely hampered by interferences stemming from polyethylene oligomers and triolein impurities (17, 18). Size exclusion chromatography can remove polyethylene oligomers and most of oleic acid and methyl oleate (18). However small amounts of oleic acid and methyl oleate remaining after size exclusion 66 chromatography are analytically problematic (18). Additionally, oleic acid in SPMD dialysates could contribute to false positive toxicity using Microtox assays (19). Petty et al (15) suggested a potassium silicate column can effectively removed oleic acid, unfortunately it can also remove target analytes (18). Gustayson et aT (17) introduced a normal phase application of a disposable column containing a dual-zone restricted-access sorbent to separate methyl oleate from polyaromatic hydrocarbons (PAHs); however this method failed to separate methyl oleate from moderately polar to polar analytes(18). Lebo et al. (18) demonstrated purification procedure for triolein destined for use in SPMD could greatly reduce interferences caused by impurities in the triolein. Nonetheless the implementation of these efforts to reduce interferences stemming from the triolein impurities would be expensive and laborious and some contaminants of interest are often sacrificed. In addition to triolein, the polyethylene membrane itself contained a significant portion of the total amount of dissolved target analytes accumulated by an SPMD in a 'aboratory aquatic system (1). Booij et al. (20) revealed the equivalent efficiency of SPMD and low density polyethylene membrane alone in sampling polychiorinated biphenyls (PCBs), PAHs, and chlorobenzenes under controlled conditions, indicating the role of triolein in the exchange kinetics was insignificant, particularly for compounds with log K0 > 6. The importance of the polyethylene membrane as a solute reservoir in a three compartment model (water, polyethylene and triolein) has been mentioned by Gale (21). Additionally, the sampling rates of SPMD and the single layered polyethylene membrane sampler were not significantly different for compounds with log K0 4-7.5 at three different temperatures (22). Unlike the triolein impurities, analytical interferences from polyethylene membrane (i.e., polyethylene oligomers or polyethylene waxes) are readily removed by size exclusion chromatography (18). Therefore preparation and sample extraction of triolein-free low density polyethylene lay flat tubing (LFT) are less laborious and less expensive. The triolein-free low density polyethylene membrane lay flat tubing (LFT) would be an alternative passive sampling device for monitoring exposure and assessment of contaminants. This simpler and less expensive sampling device would reduce the complexity of SPMD kinetic modeling and sample extraction, but provide the same benefits. However, there are some concerns for use of LFT. It was noted that the polyethylene membrane without 67 triolein appeared to be more subject to biofouling and it potentially reached equilibrium faster than SPMD because of its smaller sorption capacity (20, 21). We suspected that LFT would not reach equilibrium at ambient environmental concentrations of hydrophobic organic contaminants, suggesting the applicability of this simplified device. Also, information on the kinetic phase of chemical uptake during field deployment could be obtained by using permeability/performance reference compounds (PRCs) as an in situ calibration approach (20, 23). PRCs are analytical non-interfering compounds with moderate to relatively high fugacity from PSDs, which are added to the PSD prior to membrane enclosure(23). Because both the uptake and the dissipation of organic chemicals are controlled by the same molecular processes, in situ dissipation rates of PRCs would allow for estimating in situ PSD uptake rates (20, 23). For SPMD, triolein also served as a handy carrier for PRCs. A different approach needs to be developed for a triolein-free polyethylene membrane. Booij et al. (24) has developed spiking method based on equilibration of polyethylene membrane in 80/20 (v/v) methanol/water solution of PAHs and PCBs with log K0 3.9-7.7. However, variations in both the amount and the evaporation rates of the solvent that stuck to the membrane were sometimes problematic (24) and the membrane-solution partitioning coefficient was required. A suitable spiking method is thus needed. Although the feasibility of LFT as an alternative passive sampler in aquatic systems has been mentioned in a few studies(20-22), its field application and demonstration were very limited (25). Further study of LFT as an alternative approach for passive sampling technique is worth the effort due to its comparable efficiency to SPMD and lack of interference from the triolein impurities. Development of LFT in both theoretical and experimental work as well as field study should be explored to promote a more complete understanding of its sampling mechanism and its applicability in the field. In this study we evaluated the use of LFT as an alternative device for monitoring bioavailable hydrophobic organic contaminants in surface water and compared to to SPMD. A method for spiking PRCs into LFT was developed and their recoveries were tested in the laboratory under flowing water and in the field environment. LFT and SPMD were deployed side by side as a paired study in a contaminated harbor site in the lower Willamette River at Portland Harbor, Oregon. LFT sampling rates for selected polycyclic aromatic hydrocarbons 68 (PAHs), polychiorinated biphenyls (PCB s), and organochlorine pesticides were estimated. Field derived data demonstrated the effectiveness and reliability of LFT for environmental monitoring. Materials and Methods Passive Sampling Devices. Low density polyethylene membrane lay flat tubing (LFT) (Barefoot®) was purchased from Brentwood Plastic, Inc (St. Louis, MO, USA). The LFT was free of additives. The wall thickness of LFT was approximately 75-95 m. Standard SPMDs were purchased from Environmental Sampling Technologies (EST, St. Joseph, MO). A standard size SPMD consists of a 91-106cm segment of 2.5cm wide low density polyethylene lay flat tubing having a wall thickness of 70-95 tm and a surface area of 450 cm2, that contains 1 mL of 95% pure triolein (1,2,3-tri[cis-9-octadecenoyl]glycerol) as a thin film and has a total weight of 4.5 g (15, 16). Chemicals and solvents. Standards of organochiorine pesticides (purities and PAHs (purities PCBs (purities 98.5%) 99%) were from Chem Service, Inc. (West Chester, PA). Standard 99%) were from AccuStandard (New Haven, CT). SPMDs were fortified with PCB 82, and dibenz[a,h]anthracene for use as PRCs. List of target analytes is present in Table 3.1. Certified reference material of PCB congeners and organochlorine pesticides in cod liver oil (SRM 1588a) was purchased from the National Institute of Standards and Technology (NIST, Gaithersburg, MD). SRM 1588a was used for method validity and accuracy purposes for the deteiiiiination of PCBs and organochiorine pesticides in complex lipophilic matrices such as triolein. All solvents used were pesticide or Optima® grade from Fisher Scientific (Fairlawn, NJ). 69 Table 3.1 Selected physicochemical properties of target analytes PAHs (26, 27) chemical MW naphthalene acenaphthylene acenaphthene fluorene phenanthrene anthracene fluoranthene pyrene benz[a] anthracene chrysene benzo[a]pyrene benzo[b]fluoranthene 128 152 154 166 178 178 (NAP) (ACY) (ACE) (FLO) (PHE) (ANT) (FLA) (PYR) (BAA) (CHR) (BAP) (BBF) benzo [k}fluoranthene (BKF) (BPL) benzo[g,h,i]perylene indeno[ 1 ,2,3-c,d]pyrene (IPY) dibenz[a,h] anthracen&' (DBA) OC p, p'-DDT pesticides p ,p'-DDD (28, 29) p, p'-DDE PCBs (30, 3]) dieldrin methoxychiora endrin' - di-CBs - tn-CBs - tetra-CBs PCB 8' PCB 37 PCB 44 202 202 228 228 252 252 252 276 276 278 355 320 318 381 346 381 223 256 290 290 290 290 290 290 324 324 324 324 324 324 324 324 358 358 358 358 358 358 392 392 392 392 392 log K0 3.4 4.1 3.9 4.2 4.6 4.5 5.2 5.2 5.6 5.9 6.5 6.1 6.8 7.1 6.6 6.5 5.7 6.1 6.0 3.5 4.2 5.5 5.1 5.9 6.0 minimal box dimension (A) length breadth depth molecular volume (As) 8.9 8.8 7.2 8.4 3.1 3.1 126.9 8.8 11.1 11.5 11.7 10.7 11.4 13.7 13.6 13.6 13.6 13.3 11.5 13.2 15.6 13.1 8.1 3.2 7.2 7.7 7.2 9.0 9.5 3.1 3.1 3.1 3.1 3.1 3.1 148.8 160.4 169.5 170.3 187.7 186 13.0 13.5 10.3 13.4 10.2 9.4 7.7 8.9 9.3 9.1 10.2 10.0 9.3 8.7 9.5 8.6 8.9 11.0 4.4 8.8 8.8 3.1 4.5 3.1 3.1 3.1 3.1 212.9 212.2 228.6 230.3 231.1 244.3 255.4 8.2 7.2 5.6 7.7 6.6 234.9 259.1 PCB 49 6.1 262.8 PCB 52 6.1 11.4 7.8 5.6 262.8 PCB 60 6.3 265.2 PCB 74 6.7 4.8 13.9 7.8 PCB 77 6.5 267.6 PCB 82a - penta-CBs 6.1 PCB 87 6.5 272.4 PCB 99 6.6 PCB 101 6.4 12.6 5.6 276.1 7.8 PCB 105 6.0 PCB 114 6.7 PCB 118 7.1 PCB 126 6.9 - hexa-CBs PCB 128 7.0 282.4 PCB 138 13.9 7.8 5.6 6.7 PCB 153 6.9 13.9 7.8 5.6 289.4 PCB 156 7.2 PCB 166 7.0 PCB 169 7.6 265.2 - hepta-CBs PCB 170 7.1 PCB 180 7.2 PCB 183 7.2 13.9 8.8 5.6 PCB 187 7.2 PCB 189 7.7 a used as permeability/performance reference compounds (PRCs) in passive sampling devices molar volume (cm3/mol) 147.6 165.7 173 188 199 197 217 214 248 251 263 268.9 268.9 277 NA 300 261.3 246.4 243.1 226.4 247.3 268.2 268.2 268.2 268.2 268.2 289.1 289.1 289.1 289.1 289.1 310 310 310 310 310 330.9 330.9 70 Preparation of LFT. LFT was pre-cleaned by solvent extraction to remove impurities. Briefly, the tubing was pre-extracted twice with dichioromethane (Optima®, Fisher Scientific, Fairlawn, NJ) using a soxhlet extractor. After the second extraction, the pre-solvent extracted LFTs were dried by pulling a vacuum through polyurethane foam plugs. The apparatus was left under vacuum for approximately 48-72 hr at room temperature or until dichioromethane was removed from LFT. After dried, LFT was stored in clean, sealed paint cans, and frozen (-20°C). Pre-extracted LFTs were heat-sealed at a distance of 2 cm from one end of which had been folded to make a small loop. The fortification standard solution was pipetted into the other end of the tube. The LFT used in the laboratory experiment were fortified with 100 tL of 400 ng/mL of PCB 8, PCB 82, PCB 170 and endrin, 800 ng/mL of methoxychior, and 20 tg/mL of dibenz[a,h]anthracene in toluene. The LFT used in the field exposure were fortified with 100 L of the mixture of PCB 82 and endrin (each at 200 ng/rnL) and dibenz[a,h]anthracene (20 tg/mL) in toluene. These compounds were used as PRCs. Air was removed by squeezing the solvent down the interior of the tube toward the seal by running the tube through fingers (gloved hands). The remaining air was forced from the tube through the open end and the tube was heat-sealed. The LFT constructed in the present study was 2.7 ± 0.05 cm wide, 100 ± 0.22 cm long, with a mass of 5.5 ± 0.11 g and a surface area of 540 cm2. Fortified LFTs were individually mounted on stainless steel racks (EST, St. Joseph, MO) and stored in cleaned paint cans which were tightly sealed and stored at -20°C until deployment. Evaluation of the spiking method of permeability/performance reference compounds (PRCs) in LFT. In a pilot experiment, three fortified LFTs mounted on stainless steel racks were placed in an aquarium glass tank (35 cm x 50 m x 30 cm) filled with flowing tap water at flow rate 41 ± 7 mL/s and water temperature at 17 ± 1 °C in darkness to prevent photodegradation of dibenz[a,h]anthracene. After 14 days, LFTs were sampled and cleaned with gloved hands, then rinsed in iN HCI (for approximately 15 sec) (Trace metal grade, Fisher Chemical, Fairlawn, NJ), 18 MQcm water, acetone (Pesticide grade, Fisher Scientific, Fairlawn, NJ), and isopropanol (Pesticide grade, Fisher Scientific, Fairlawn, NJ), respectively. LFTs were kept in clean glass amber jars and stored at -20 °C until analysis. 71 Unexposed fortified LFT, fortified LFT procedural blank, reagent blank, and fortified reagent blank accompanied the exposed LFTs and were processed and analyzed exactly as the exposed samples. Demonstration of effectiveness and reliability of LFT for field deployment. LFTs were tested at a field site in the lower Willamette River at Portland Harbor which is listed as an impaired water body in the Federal National Priority List (NPL, so called a superfund site) (32). The harbor sediments have been contaminated with PCBs, DDTs, PAHs, dioxinlfurans, and heavy metals (33). Fish from this area were contaminated with DDTs and PCBs (34). The Willamette River at Portland Harbor is deep, slow moving, and tidally influenced (33). The harbor generally has a low sediment transport capacity (33). Surface water runoff is determined by climate influence and varies according to season (33). LFTs were deployed side by side with SPMDs at the lower Willamette River at Portland Harbor from River Mile 1 to 18.5 which contained the Portland Harbor superfund site (RM 3.5 to 9.5) and the McCormick and Baxter Creosoting Co. superfund site (RM 7 East). The eleven sampling sites were outside the main shipping channel and often near stream outlets of various upstream land uses including industrial, urban/residential, and undeveloped areas. Five individual PSDs were loaded in a flow-through stainless steel cage. Duplicate cages of LFTs and SPMDs were submerged approximately 10 ft from the river bottom and were suspended with an "anchor-cable-cage-cable-float" arrangement. PSDs were deployed for 21 days. The average water temperature ranged from 9 to 22 °C. The five PSDs were later combined for analysis. On retrieval, PSDs were cleaned by gently rubbing with gloved hands in on-site water and the series of solvents described above. Cleaned PSDs were kept in clean glass amber jars and transported on ice-packs. Samples were stored at 20°C until analysis. Sample extraction and chemical analysis. PSD extraction and cleanup were based on established protocols and only modified slightly as necessary (5, 15). Briefly, PSDs were dialyzed in hexanes, 400 mL for 5 PSDs (Pesticide grade, Fisher Scientific, Fairlawn, NJ) for 18 hr, followed by a second dialysis with fresh hexanes for 6 hr. The combined dialysates were concentrated by rotary evaporation and a TurboVap® LV evaporator (Zymark®, 72 Hopkinton, MA). The samples were cleaned and fractionated using gel permeation chromatography (Waters® gel permeation chromatography cleanup system; Water® 515 HPLC pump, 717 Plus autosampler, Phenogel® column, 2487 dual X absorbance detector, and fraction collector II) (Milford, MA) with dichloromethane (Opima®, Fisher Scientific, Fairlawn, NJ) as the mobile phase at the flow rate of 5 mLlmin. Appropriate fractions were determined by analyzing standards and fortified samples. The appropriate fraction (14-20 mm) was collected and split into two equal sub-fractions: 1) PCB and OC pesticides and 2) PAHs. Both fractions were separately subjected to volume reduction using a TurboVap® LV evaporator and solvent exchanged into iso-octane (Pesticide grade, Fisher Scientific, Fairlawn, NJ) for PCB and OC fraction and acetonitrile (HPLC grade, Fisher Scientific, Fairlawn, NJ) for PAH fraction with a final volume of 1 mL. PCB congeners and OC pesticides were deteuiiined using GC-tECD (Agilent Technologies 6890N Network GC system) (Palo Alto, CA) dual capillary columns (db-xlb and db-17, J&W Scientific Inc., Agilent Technologies, Palo Alto, CA) /dual detectors. Injector and detector temperature were at 250 °C, and 350 °C, respectively. The GC system was operated with helium carrier gas and nitrogen makeup gas. The oven temperature program was as follows: 100 °C (1 mm) and increased at 1.2 °Cmin1 to 265 °C (2 mm). PAH detection and quantitation was performed on a Hewlett Packard HPLC series 1100 (Agilent Technologies, Palo Alto, CA) with dual detection by fluorescence and diode array, both with multiple wavelengths. The fluorescence detector had an excitation wavelength at 230 and emission wavelengths at 360, 410, and 460 nm; the diode array had detection signals at 254, 242, and 230 nm. Only three compounds, fluorene, acenaphthylene, and indenol (1,2,3-c,d) pyrene, were detected by diode array; the rest were detected with the fluorescence detector. The column was a Phenomenex Luna C18 with 3-tm particle size (Phenomenex, Torrance, CA). The instrument was run with a constant flow rate of 0.75 mL/min and a timed gradient for the acetonitrile/water eluent system. The time program ran at 40% acetonitrile for 10 mm, was gradually ramped up to 70% acetonitrile for 15 mm, and then ramped up to 90% acetonitrile for 19 mm. The program was held at 90% acetonitrile for 3 mm and then returned to 40%. 73 Field duplicates and field blanks (trip blank, field blanks, and field extraction blanks) accompanying the deployed passive sampling devices during deployment, retrieval, and transportation to the laboratory were included in every batch. These field blanks and laboratory control blanks (i.e., procedural blanks, laboratory PSD controls, fortified samples, and standard reference material) represented 30 to 40% of a sample set and were processed and analyzed exactly as the deployed samples. The method detection limits (MDL) were determined as three times the heights of coincident peaks observed for each compound in the laboratory PSD control. For those analytes having no coincident peak, the MDLs were set at a value equivalent to the lowest standard in the respective calibration curve. None of the target analytes were identified in field blanks and laboratory control blanks. The average percent recoveries of fortified samples were as follows: PAHs 70 ± 20%, PCBs 66 ± 13%, p, p'-DDT 99 ± 20%, p, p'-DDD 85 ± 8%, p, p'-DDE 79 ± 7%, and dieldrin 75 ± 17%. The average percent recoveries in standard reference material were as follows: PCBs 96 ± 24%, p, p'-DDT 99 ± 21%, p, p'-DDD 108 ±14%, p, p'-DDE 79 ± 11% and dieldrin 114±27%. Data analysis. Data interpretation was performed using SPSS® Version 10.0.1(SPSS Inc., 1989-1999), Sigma Plot 2002 for Windows Version 8.0 (SPSS Inc., 1986-2001), and Microsoft® Office Excel 2003 (Microsoft Corporation, 1985-2003). Standard descriptive statistics, paired t-test, linear regression techniques were used. In the field exposure experiment, the efficiency of LFT to sequester hydrophobic contaminants was evaluated. The efficiency of two PSDs was analyzed by comparing amounts of contaminants taken up per unit mass of LFT and SPMD using paired samples t-test. Relationships of LFT concentrations, SPMD concentrations, and physico-chemical parameters for PAHs, PCBs, and organochlorine pesticides were examined by linear regression approach. Results and Discussion Evaluation of the spiking method of permeability/performance reference compounds (PRCs) in LFT. In the laboratory study with LFT, LFTs were spiked with 74 selected PAHs, PCBs, and organochiorine pesticides in a small volume of toluene (100 tL). These compounds were used as PRCs (Table 3.2). Averaged recoveries of PRCs in unexposed LFTs were 75% except only for PCB 8 (2, 4'-dichlorobiphenyls) (log K0 = 5.1) which yielded an averaged recovery of 62% (Table 3.2). PCB 8, a two chlorine substituted congener, has a relatively high vapor pressure (6.9 x 102 Pa) (30) compared to other target contaminants. It is plausible that volatilization of PCB 8 during sample extraction and analysis could contribute to its loss. In general, the percentage recoveries of unexposed LFT procedural control samples 75% indicated the dissipation of spiked compounds during sample preparation, storage and handling, and extraction was negligible. The spiked compound integrity was maintained until analysis. In addition, a stability test of spiked LFTs that were kept in closed amber jars at room temperature (22-25°C) for three weeks yielded an average recovery rate of 89% ± 11%. No significant loss of spiked compounds in the stability test suggested loss of sample integrity during transport or storage under 22-25°C would be negligible. Table 3.2 Percent recoveries of permeability/perfoj titance reference compounds using simple spiking method in a small volume of toluene. permeability/performance reference compound (PRC) PCB 8 (log = 5.1) PCB 82 (log = 6.1) PCB 170 7.1) (log K0 endrin 5.5) (log K0 methoxychior (log K0 = 4.2) dibenz Ia,hj anthracene (log K0= 6.5) a =2 percent recovery (%) exposure under unexposed laboratory flowing water (n=3) control (n=9) PRC clearance rate keprc (d') 26±8.1 62±11 0.06 67±2.5 75±9.1 0.008 86±4.0 87± 10 0.0008 98±7.1 110±20 0.008 53±16 114±15' 0.05 91±4.2 93±13a 0.002 75 According to PRC approach, measuring the PRC loss over the exposure period provides an in situ clearance rate constant of PRC (keprc) (23), keprc = in (CpsDO / CPSD) t' (1) where CPSDO is the initial concentration of the PRC and CPSD is the concentration of PRC remaining in the PSD following exposure, and t is exposure time in days. Comparison of the keprcfield derived from the field exposed PSD to the keprclab measured in calibration exposure (keprcfield / keprclab) can serve as an indicator of differences in the exposure conditions or the effect of environmental variables on PSD sampling (23). In a two-week laboratory flowing water exposure study, keprclab for compounds with log 5.5 were less than 0.008 d' (Table 3.2). This finding confirms that polyethylene membrane without lipid reservoir (i.e., triolein) has the capacity to retain hydrophobic organic compounds by diffusive process in the polymer (21). The simple spiking method of PRCs in a small volume of toluene was effective and valuable for further development of LFT application. The result also suggests spiking with a small volume of other non-aqueous organic solvents such as iso-octane is applicable. This method is simple, inexpensive and provides the same benefit of a permeability! performance reference compound approach. Demonstration of effectiveness and reliability of LFT for field deployment. As part of our ongoing project for evaluation of seasonal bioavailability of contaminants in surface water at the lower Willamette River at Portland Harbor, Oregon (35), passive sampling devices were deployed for a period of 21 days. LFTs and SPMDs were deployed side by side as the paired study at thirteen sampling sites in three different conditions; average water temperature 9 ± 1°C /average river flow 14000 ± 4800 ft3/s, average water temperature 22 ± 1°C !average river flow 8300 ± 390 ft3/s, and average water temperature 22 ± 0.3 °C /average river flow 10000 ± 1700 ft3/s. The total of 74 composite samples (each sample was a composite of 5 PSDs) of LFT and SPMD were compared. PCB 82, endrin and diben4a,hjanthracene were used as PRCs in both LFTs and SPMDs to account for the effects of environmental conditions on the device uptake rate. 76 Evaluation of LFT following field deployment. The outer surface of the body of LFTs and SPMDs, following each deployment period, was covered in a thin film of silt. It appeared that the samplers deployed under condition of water temperature 22°C I river flow 8300 ft3/s in summer had slightly more fouling as compared to the other two conditions. However, in all cases the degrees of fouling on the exterior surface of LFTs and SPMDs which were deployed for 21 days were not significantly noticeably different. This observation differs from previous observations in which triolein-free SPMD was reported to be more subject to biofouling than SPMD (20, 21). After 21-day deployment of each sampling event, the total of 185 LFTs were not observed to have more biofouling than SPMD. The membranes of LFTs and SPMDs appeared to be free from biofouling and algae growth. Heavy biofouling was reported to reduce the permeability of diffusive-limiting membrane of SPMDs, resulting in lowering the sampling rate by as much as 30% for phenanthrene (log K0 = 4.6) (36). Additionally, it was noticed that the membranes of LFIs after deployment were slightly more rigid than the membrane of SPMDs. The lack of rigidity of triolein-containing SPMD is plausibly due, in part, to the formation of a thin film of triolein derivatives or triolein impurities on the membrane exterior surface which might affect the contaminant uptake mechanism of SPMD. Evaluation of the influence offleld condition on contaminant uptake kinetic of LFT. Both uptake rate and clearance rate of hydrophobic contaminants in the passive sampler are governed by the same molecular processes (21). It has been proposed that any change in the uptake rate of contaminants to the passive sampler should be reflected by a change in the clearance rate from the passive sampler (20, 23). The device uptake rates may change due to changes in temperature, flow velocity of the surrounding water, and biofouling on the membrane surface during field exposure (23). The use of PRCs as an independent measure for the exchange kinetics between the passive samplers and water allows for device uptake rate adjustment for these field variables (20, 23). The ratio of the clearance rate constant (keprcfield) in the field and in the laboratory (keprclab) can serve as an indicator to differences in the environmental conditions. In addition, the information on the kinetic phase of contaminant uptake under field exposure can be obtained from the clearance of PRCs in the field (20). It has been suggested that the exchange kinetics are under aqueous boundary layer control for compounds with log K04.4-8.0 and under polyethylene membrane control for 77 compounds with log K0 < 4.4 (23). However, it has been proposed that the use of PRCs with log K0 4.4-5.5 was also applicable to the target analytes under membrane control (23). Thus, the range of physico-chemical properties of PRCs used in this study was representative of all target analytes. The PRC approach was used in both LFTs and SPMDs in the present study. Table 3.3 presents percent recoveries and clearance rates (keprcfield) of PRCs in LFTs and SPMDs which were paired-deployed under two different conditions. The PRC comparison study was done at 22 °C. Booij et al. (24) suggested complete dissipation of a PRC with log K0 of 5 indicating all analytes with similar and lower log K0 had attained equilibrium phase. In contrast, no dissipation of a PRC with log K0 of 6 indicating all analytes with similar and higher log K0 were in the linear uptake phase (24). Significant retention of PRCs with log K0 of 5.5 to 6.5 in both LFTs and SPMDs (Table 3.3) suggested linear uptake phase for all target analytes could be assumed. Small deviation of the PRC recovery rates within each condition indicated insignificant variation from analyses or site-specific condition variation among sampling sites. Contaminant clearance times by LFT and SPMD were similar. The mean PRC clearance rate (keprcfield) in LFT and SPMD was not different (0.02 d1 for LET and SPMD at water temperature 22 °C, river flow 8300 ft3/s; and 0.02 d' for LFT and 0.01 d' for SPMD at water temperature 22 °C, river flow 10000 ft3/s). Similar clearance time indicated for a 21-day field exposure for both LFT and SPMD were governed by the same exchange kinetics. PCB 82, endrin, and dibenz {a,h] anthracene clearance rates in the LFTs were not different between the two conditions (Table 3.3A and 3.3B). In contrast, the clearance rates for dibenz [a,h] anthracene in SPMDs were somewhat different between the two conditions. This finding suggests the exchange kinetics controlling more hydrophobic compounds such as dibenz {a,h] anthracene (log K0 = 6.5) in LFTs is more robust to the changes in environmental condition than in SPMDs. 78 Table 3.3 Percent recoveries and clearance rates of permeability/performance reference compounds in the field exposed polyethylene lay flat tubing (LFT) and semipermeable membrane device (SPMD) under different field conditions. Average water temperature 22°C, average river flow 8300 ft3/s, percent recovery (%) field sample laboratory and field (n131 control (n=7) LFT SPMD LFT SPMD permeability/performan ce reference compound (PRC) PCB 82 6.1) (log K0 endrin (log 5.1) PRC clearance rate kep (d1) keprc.field/ keprclab LFT SPMD LFT 62±2.2 50±3.0 81 ± 8.2 80 ± 5.9 0.01 0.02 1.25 41±4.9 64±4.9 93±24 80±10 0.04 0.01 5 92±7.2 94±4.2 0.004 0.03 2 dibenz{a,hJ anthracene 84 ± 6.2 52± 13 6.5) (log K0 a each sample was a composite of 5 PSD Average water temperature 22°C, average river flow 10000 ft3/s. permeability/performance reference compound (PRC) PCB 82 (log K0 = 6.1) endrin 5.1) (log K0 dibenz[a,h] anthracene (log K0= 6.5) percent recovery (%) field sample laboratory and field (n=12y control (n=7) LFT SPMD LFT SPMD PRC clearance rate keprcfield/ keprc (d') kepc4ab LFT SPMD LFT 65±7.8 54±6.0 73 ± 3.4 85 ± 8.9 0.006 0.02 0.75 33±7.7 89± 11 87±4.7 106± 15 0.04 0.008 5 85±4.4 65±7.6 92±8.5 100±4.8 0.004 0.002 2 a each sample was a composite of 5 PSD Clearance rate ratios of PRCs between field and laboratory exposure in LFTs were not different either of the conditions tested (Table 3.3A and 3.3B). This suggested the increase of river flow from 8300 ft3/s to 10000 ft3/s or 20% difference did not significantly affect on the contaminant uptake kinetics of LETs. The similar clearance rates of PRCs from Table 3.3A and 3.3B indicated the biofouling effect and river flow variation between the two conditions were negligible. Although data for PRCs were not available for the sampling event of water temperature 9°C, a temperature effect can be expected (22, 27, 29, 37). The contaminant 79 uptake rate by polyethylene membrane is dependent on temperature in both aqueous boundary phase control and membrane phase control (16). Temperature can affect molecular diffusivity (38). Elevated temperatures result in more Brownian movement and elevated temperature media are less densely packed so that diffusivity of chemicals through them is facilitated. Evidence of the temperature effect on polyethylene membrane uptake rates has been documented (22, 27, 29, 37). For compounds under aqueous boundary control (log K0 4.4) (22) , the effect of a 13°C temperature change (from 9°C to 22°C) on aqueous diffusivity can be estimated from the ratio of diffusion coefficient in water (D) at 9°C and 22°C from the Stroke-Einstein relation and the equation by Hayduk and Laudie (38). A 1.5- fold increase in D was computed for a 13°C increase in temperature (D @ 22°C! D @ 9°C = 1.5). It is noted that this calculation could not apply to the membrane boundary layer controls (i.e., polyethylene membrane). However, temperature increase in the chemical diffusivity in polyethylene membrane can be expected as there is increased polymer chain movement within the membrane due to an elevated temperature. Any temperature increase in the aqueous diffusivity and the membrane diffusivity should cause a corresponding increase in the uptake rate by LFT. Because contaminant partitioning and diffusion occurred at both aqueous boundary phase and polyethylene membrane layer, the uptake rates for compound with log K0 4.4 are expected to increase at least 1.5 fold when water temperature changes from 9°C to 22°C. Comparison of contaminant accumulation in LET and SPMD. The paired LFT and SPMD field exposure study demonstrated that LFT accumulated PAHs, PCBs, and organochiorine pesticides as effectively as SPMD (Figure 3.1). Linear correlation between contaminants concentrations in LFT and in SPMD (amounts of contaminants accumulated per mass of PSD) (ng/g) indicated both devices functioned similarly. The capacity of LFT and SPMD to accumulate PAHs, PCBs, and organochlorine pesticides was approximately equivalent as expected from their similar surface areas. Lack of triolein as an ultimate reservoir in the device did not affect the capacity of LFT to accumulate hydrophobic organic contaminants under realistic exposure conditions. It has been hypothesized that a polyethylene membrane-only sampler reached equilibrium faster than triolein filled sampler (i.e., SPMD) because of their smaller sorption capacity, making it a poor choice for integrative long term monitoring (21, 22). Our study was conducted at a superfund site where 80 the river system has been contaminated with a wide range of contaminants. The result from our study indicated that LFT did not reach equilibrium state during a 21-day field exposure at a highly contaminated system. This finding indicated LET performed at least as well as SPMD within the environmentally observed ranges of PAHs, PCBs, and organochiorine pesticides. Concentrations of contaminants accumulated in LFT and SPMD at each sampling site for each deployment were statistically compared (a paired t-test, two-sided p-value 0.05 was considered significantly different). The mean concentrations for p,p'-DDT, p,p'-DDD, p,p'-DDE, and PCBs (sum of 25 individual PCB congeners) in LFT were higher than in SPMD in all three different conditions. The congeners with log K0 ranging from 6.0 to 7.0 were more concentrated in LEE than in SPMD. No statistical differences were observed between the concentrations in LEE and in SPMD for the congeners with log K0 < 6.0 or> 7.0. The mean concentration of dieldrin, by contrast, was higher in the SPMD. No significant difference was observed for the mean concentration for PAHs (sum of 15 priority pollutant PAHs). However, the mean concentrations of three-ring PAHs with log K0 <4,5 were higher in the SPMD. On the other hand, the mean concentrations of PAHs with log K0 6.0 were higher in LFT. No differences were observed between the concentrations in LEE and in SPMD for PAHs with log K0 4.5-5.5. 81 4 V V .-0 01 50.8- 0 0 0.6 -5 0.4 ci) 0.2 r2=0.9331 = 0.7255 J) < 0.0001 p<O.0001 0.0 00 0.2 0.6 0.4 0.8 12 1.0 0 dieldrin in LFT (ng/g) 25 20 0 15 5 6 3.5 a) .E. 3.0 0 V 0 2.5 0 10 C/) 2.0 - C C 0 0 4 4.0 0 C Cl) 3 2 1 pp-DOT in LFT (nglg) S w 1.5 0 1.0 a- - =0.9488 p<O.000l 0.0 0 5 15 10 25 20 p,p-DDD in LFT (ng/g) 30 0 1 2 3 4 5 6 7 pp-ODE in LFT (ng/g) 3000 -2500 a) 2000 1500 C/) . 1000 - 50:: 0 2 4 6 FOBs in LFT (nglg) 8 10 0 500 1000 1500 2000 2500 3000 PAHs in LFT (ng/g) water temperature 9°C, river flow 14000 ft3ls o water temperature 22°C, river flow 8300 ft3/s water temperature 22°C, river flow 10000 ft3/s Figure 3.1 Relationship of contaminant concentrations in the triolein-free polyethylene lay flat tubing (LFT) and in semipermeable membrane device (SPMD). Linear correlation suggests both devices are governed by the same uptake model and function similarly. This indicates LFT can accumulate the selected contaminants as effectively as SPMD regardless of field variations. 82 Spatial distributions of contaminants accumulated in LFT were compared to those in SPMD. Figure 3.2 is an example of three deployments during 2003 to 2004. This 21-day exposure was taken during a period of water temperature 22 °C, river flow 8300 ft3/s in summer of 2004. The spatial patterns for organochiorine pesticides and PAHs in polyethylene lay flat tubing were identical to the patterns in SPMD. The spatial patterns for PCBs in the two passive samplers were only slightly different at some sampling sites. This is mostly likely due to SPMD PCB concentrations near detection limits. A small variation between the PCB distributions in LFT and in SPMD at those sites is less significant especially when one considers the close resemblance of contaminants at the other sites. Compositions of PAHs and PCBs accumulated by LFT and SPMD from side by side field deployment were also very similar (Figure 3.3). The field exposure demonstrated the congener distributions and total concentrations were strongly correlated between the two sampling devices. The LFT/SPMD concentration ratio was independent of temperature and flow regime in this study. The LFT/SPMD concentration ratio versus log K0 (Figure 3.4) shows that for the forty-five contaminants tested, twenty-three were found to uptake in LFT faster than in SPMD (below detection limit contaminants were excluded from the discussion). For contaminants with log K0> 5.0, except PCB 37 and PCB 187, the LFT/SPMD concentration ratio is 1. Only ten compounds, generally all with log K0 4.5, were found to have LFT/SPMD concentration ratios < 1.0. This finding indicates the role of triolein is insignificant for a compound with log K0 5. For a compound with less hydrophobicity, triolein had more effect on contaminant accumulation. The effect of triolein on the amount taken up was about a factor of two for a compound with log K0 4.5. In a laboratory flow control study by Booij et al. (20) using four SPMDs without triolein, the ratios of amounts taken up by SPMD without triolein / with triolein were equal for compounds with log K0> 6, and the effect of triolein on the uptake kinetics was as large as a factor of four for a compound with log K0 = 4. Our field study results corroborated the laboratory exposure results by Booij (20), indicating the overall effectiveness and reproducibility of LFT for sampling hydrophobic contaminants. 83 0) 0) dieldrin 25 c 25 - -0-- p,p- DDE -'- p,p-DDD v--- p,p-DDT 10 20a. 15 ci) 10. C 4C) 0 .2 5 .:: 0- 0- C C) C 8 b0 0) 10 £8 PCBs C) £ 8- F- 0 C (I) U- ._i6 0 4 . C 02 C) CC C) Co 0 C.) C 0 C C)0 0 S 0) 3000 2500 - PAHs 0 0 2000 ci C 0 S 2500 0 2000 3000 0) 1500 - 0)1500 1000- C 1000 .2 500- 500 0.. C) 0 b <D < o Figure 3.2 Comparison of spatial distributions of selected contaminants accumulated in the triolein-free polyethylene lay flat tubing (LFT) and in semipermeable membrane device (SPMD). LFT and SPMD were deployed side by side at the lower Willamette River at Portland Harbor, Oregon for 21 days in summer of 2004. Duplicated samples were deployed at River Mile 1 and River Mile 18. 20 LFT PCBs = 5.61 ± 0.31 nglg n=2, site = RM 1 22°C, 8300 ft3Is 0 0 0 10 84 60 I 50 LFT PAHs = 523 ± 59 ng/g 40 n=2, site = RM 1 9°C, 14000 ft3/s -1 30 ci, 0 0 20 10 1) ci) .20 SPMD PCBs = 4.12 ± 0.30 ng/g n=2, site = RM 1 ci, 0 22°C 8300 ft3/s 10 0 60 SPMD I< 40ci) PAHs=618+210ng/g 50- n=2, site = RM 1 9°C, 14000 ft3!s a- -1 30- 0 0 N C) C'JO r- NC)- Lfl OuJF<tI<iuJ)< <OOJiz-i>-<I<QJQ-OaJ C')(D CD C)OQC) N. C) QOQQftQOQOOOOOOOCOOOOOOOO OQOOO. Figure 3.3 Percent composition of PCBs and PAHs (average ± lsd) in the triolein-free polyethylene lay flat tubing (LFT) and semipermeable membrane device (SPMD) which were paired-deployed at the lower Willamette River at Portland Harbor, Oregon for 21 days. Total concentrations and field condition are noted in each panel. See Table 1 for abbreviations. 0.0 3 4 5 6 7 8 log K0 Figure 3.4 Average ratios of concentration of contaminants accumulated in the triolein-free polyethylene lay flat tubing (LFT) and in semipermeable membrane device (SPMD) as a function of octanol-water partitioning coefficient (log K0). See table 3.1 for abbreviation. 85 Estimation of LET contaminant sampling rates. The present study demonstrates LFT is as efficient in sampling hydrophobic organic contaminants in water as is SPMD. The efficiency and reliability of LFT suggest the neutral lipid reservoir (i.e., triolein) is not always necessary. The LFT passive sampling device is theoretically less complex than SPMD. LFT can be modeled as a two compartment system while SPMD is a more complicated three-compartment system (21) Although less complex, the LFT passive sampling device maintains significant capacity to sample waterborne contaminants in a polluted eco-system. However, field application of LFT limited due to the lack of daily sampling rates for contaminants. The contaminant sampling rates allow back-calculation of aqueous contaminant concentrations. The input to the lack of established sampling rates for a wide array of contaminants is necessary to provide assessment to water quality standard, as well as temporal and spatial comparison. The results in the present study, as well as, those by others (21, 22) indicate LFT or polyethylene membrane alone share the same contaminant uptake mechanisms with SPMD. Dissolved hydrophobic contaminants in water are transported by diffusion in the bulk aqueous phase to the aqueous diffusion film, then diffusion through the aqueous film and partition into the polyethylene membrane (21). Booij et aL (22) observed the difference between polyethylene membrane-water partitioning coefficients and SPMD-water partitioning coefficients from fifteen samples at three different temperatures were less than 0.6 log unit in the 4< log K0 <6, suggesting the significant role of polyethylene membrane in contaminant uptake. There was no evidence of significant differences observed between sampling rates of SPMD and single-layered polyethylene membrane sampler in their study (22). The percent recoveries and clearance rates of PRC indicated the linear uptake model for LFT samplers and SPMDs deployed in this study. Estimated water concentration (C) by polyethylene containing passive sampling device (PSD) in the linear uptake model is obtained from the following equation (16) cw = CPSD MPSD R' t - (2) 86 where CPSD is the concentration of the contaminant in the PSD (i.e., SPMD, LFT), MPSD is the mass of PSD in grams, R is the sampling rate in liters per day of PSD, and t is the exposure time in days. In the present study, LFT sampling rates for PAHs, PCBs, and organochiorine pesticides were estimated as a function of SPMD sampling rates under the same field conditions. When LFTs and SPMDs are pair-deployed, such as in this study, water concentration and other variables are the same for the paired deployment, leaving an R as the only unknown variable. Established SPMD sampling rates for PAHs, PCBs, and organochiorine pesticides at multiple temperatures have been well documented (27, 29, 37, 39). LFT' sampling rates can be theoretically estimated from calibrated SPMD sampling rate. RS.LFT = CLFT. MLFT. R r-SPMD CSPMD MSPMD' (3) Table 3.4 presents the LFT sampling rate estimates (R) at 9 °C and 22 °C. The LFT sampling rates were found to increase with increasing log K0 and exposure temperature (Table 3.4 and Figure 3.5) as has been observed in SPMD (22). Large variation of LET sampling rates was observed as log K0 and molecular size increased (Figure 3.5). Membrane permeability limitation could be a factor contributing to this variation. Similar to biomembrane cavities, the diameters of polyethylene membrane transient cavities range up to 9-10 A (15). This cavity size limitation impedes the diffusion of compounds with large molecular size or particle-bound contaminants through the polyethylene membrane. Contaminant molecules having at least two dimensions less than 10 A can readily diffuse through the membrane cavities. Due to membrane cavity size limitation, reduction of LET sampling rates for high K0 compounds with breadths A, such as benz[a]anthracene and benzo[k]fluoranthene, were observed (Table 1 and 4, and Figure 5). On the other hand, the breadths of high K0 PCB molecules are < 9 A (Table 1) which allows them to diffuse through the membrane cavities. Also, the biphenyl bond of PCBs allows molecular conformation and rotation which may increase their membrane permeability. These differences in compound characteristics may contribute to large variation of LFT' sampling ratçs for high K0 compounds. Elevated temperatures may decrease the variation of LET 87 sampling rates for high K0 compound by increasing polymer chain movement within the membrane, which can facilitate chemical transport through the membrane. Table 3.4 Contaminant sampling rates for triolein-free polyethylene lay flat tubing (LFT) estimated from field exposure to contaminated water at the lower Willamette River at Portland Harbor, Oregon for 21 days. LFT sampling rates were estimated from established SPMD sampling rates. compou nd class PAils compound naphthalene acenaphthylene acenaphthene fluorene phenanthrene anthracene fluoranthene pyrene benzLa]anthracene chrysene benzo[a]pyrene benzo[b]fluoranthene benzo[k]fluoranthene OC p,p'-DDT pesticides p,p'-DDD p,p'-DDE molar volume (cm fmol) 147.6 165.7 173 188 199 197 217 214 248 251 263 268.9 268.9 261.3 246.4 243.1 dieldrin PCBs -tn-CBs PCB 37 PCB 44 PCB 49 PCB 52 PCB 74 247.3 268.2 268.2 268.2 -penta-CBs PCB 87 289.1 289.1 289.1 289.1 310 310 310 - tetra-CBs PCB 101 PCB 105 PCB 118 -hexa-CBs PCB 138 PCB 153 PCB 156 - hepta-CBs PCB 170 PCB 180 log K0. 3.4 4.1 sampling rate, Lid a 9C n 1.3 ± 0.8 2.2 ± 0,8 11 39 b 4.2 4.6 4.5 5.2 5.2 5.6 3.4 ± 6.5 6.1 6.8 5.7 6.1 6.0 3.5 5.9 6.0 6.1 6.1 22 C R 12 b 1.0 4.3 ± 2.0 3.5 ± 0.5 3.7 ± 0.4 4.5 ± 0.5 1.9 ± 0.3 12 12 11 12 12 2 b b b b b b 2.4 ± 0.6 1.5 ± 0.5 2.3 ± 0.6 3.5 ± 0.8 4.2 ± 0.9 6.9 ± 1.1 8.3 ± 0.9 6,4 ± 1.3 4 13.7 ± 2.1 9.5 ± 2.6 5.8 1 b b 7.1 4.1 ± 0.5 7.3 ± 1.4 6 4 3.5 ± 5 7.4 ± 1.3 1.0 6 b b b b 8 22 21 10 12 7 4.5 ± 0.8 7.1 ± 4.7 6.3 ± 0.9 7 4.6 ± 1.4 7.7 ± 3.8 7.9 ± 2.6 8.7 ± 1.8 b 1.5 1.1 1.3 13 6 13 b 9.7 ± 6.0 ± 5.9 ± b 12.6 ± 4.9 12.9 ± 6.4 3.5 ± 1.1 2.4 ± 0.3 1.6 ± 0.3 1.1 ± 0,1 6.7 6.5 6.4 6.0 7.1 b 11 12 12 12 12 7 12 3 6.7 6.9 7.2 fl b 13.6 ± 3.0 11.5 ± 3.6 17.1 ± 3.3 16.8 ± 2.9 17.3 ± 3.9 11.5 ± 2.0 19.7 ± 3.0 16.6 ± 8.6 15.0 ± 4.2 12.4 ± 3.8 10.0 ± 3.1 16.5 ± 1.8 13.9 ± 4.7 7.4 ± 1.7 20 24 25 25 25 10 25 24 25 10 15 19 23 2 8 25 5 8 b b 7.2 6 b b PCB 183 4 7.2 b b PCB 187 330.9 7.2 14 values are average sampling rates; number in parenthesis are I SD a sampling rate estimates for 1 triolein-free polyethylene lay flat tubing (LFT) sampler with a 100 cm long and 2.7 cm wide with a mass of 5.5 g and a surface area of 540 cm2 b no value given due to concentration below detection limits 330.9 88 100 o 100 temperature 9 °C temperature 22 °C temperature 9 °C temperature 22 °C 0 10 0 3 4 5 6 8 7 100 120 140 160 180 200 220 240 260 280 300 log K0 molecular volume (A3) 100 100 o temperature 9°C temperature 22°C o temperature 9°C temperature 22 °C Oo 0 00 S U- 0 10 -1 S S 100 150 200 250 300 molar volume (cm3/mole) 350 20 S 0 I 40 80 60 minimal molecular two-dimensions (A2) Figure 3.5 Sampling rate estimates (R2L) of a 540 cm2 triolein-free polyethylene lay flat tubing (LFT) as a function of log K0 ,molecular volume, molar volume, and minimal molecular two-dimension. From Figure 3.5, the initial sharp increase of sampling rates with increasing log K0 and molecular size is indicative of membrane-controlled uptake while the relative constancy of sampling rates at the large log K0 and molecular or molar volume is indicative of aqueous boundary layer-controlled uptake. For membrane-controlled uptake, R is proportional to the product of the mass transfer coefficient for the polyethylene membrane (k1) and the polyethylene membrane-water partition coefficient (K) (22). Increasing molecular size and shape parameters can reduce membrane diffusivity (40). By contrast, since Kmw 5 hydrophobicity-related, it can be expected that increase in molecular weight, which is proportional to molar volume and log K0, would increase Kmw. For aqueous boundary layer- controlled uptake, the mass transfer coefficient for the aqueous boundary layer is a function of the kinetic viscosity of the water, and the compound's aqueous diffusion coefficient (D), 89 which is negatively related to molecular size (40). The difference in LFT sampling rates between compounds under aqueous boundary layer control can be preliminarily predicted from water diffusion coefficient (D). However, it was found that aqueous diffusivity alone cannot explain the difference in LFT sampling rates between compounds. For instance, within the same PCB homolog group which has equal chlorine numbers substituted and equal molar volume, LFT sampling rates for congeners with higher log K0 tended to have higher LFT sampling rates. The result suggests although molecular size is a major factor controlling aqueous diffusivity for a compound under the aqueous boundary layer-controlled uptake, the hydrophobicity (log K0) that controls the chemical partitioning in the polyethylene membrane maintains a significant role in contaminant uptake by the LFT sampler and cannot be neglected. According to the effect of aqueous diffusivity, a 1.5-fold increase in LFT sampling rates for compounds under aqueous boundary layer control (log K0 4.4) was expected when water temperatures changed from 9°C to 22°C (see section Evaluation of the influence offield condition on contaminant uptake kinetic of LET). The estimated LFT sampling rates for compounds with log K0 ranging from 4.4 to 7.2 rose an average of 3.4-fold with the 13 °C temperature increase. The difference between the predicted and field exposure values indicated the temperature effect on aqueous diffusivity of a contaminant contributed 40% to an increase in LFT sampling rate for a temperature difference of 13 °C. This suggested that the other parameters involved in the uptake kinetic processes such as in the polyethylene membrane boundary layer (i.e., membrane diffusivity, log K0, and molecular size) are also significant contributors to an increase in LFT sampling rates for compounds under aqueous boundary layer control (log K0 4.4). Combining the effects of aqueous diffusivity, log K0, and molecular size on the LFT sampling rates, the LFT sampling rates can be modeled using a linear regression model, see Figure 3.6. Organochiorines and PAHs were separately modeled. Both models were statistically significant (p <0.05), suggesting the LFT sampling rates can be predicted from the product of the aqueous diffusivity, log K0, and the minimum molecular two-dimensions of a contaminant. The fitting models for these two contaminant classes confirm the important roles of chemical diffusion in both polyethylene membrane (log K0, and minimal 2- 90 dimension) and aqueous boundary layer (aqueous diffusivity, molar volume). The LFT uptake rates for other PCB congeners or PAHs can possibly estimated using equations in Figure 3.6. 20 y =37.99x-6.95 P r2 = 0.79 101 138 15 - p <0.001 .PCP. 153 p,p4D C152 10p,p DD PR B 5- p,pOT pp-SaD: PR .PC153 ,p4DE P F ANT A 0- y =7.15x-0.71 r2 = 0.27 p =0.028 -5 00 D 0.2 0.4 0.6 0.8 10 1 O6.Cm2.S). Log K0Jminjmaj molecular 2-dimension (A2) Figure 3.6 Sampling rate estimates of a 540 cm2 triolein-free polyethylene lay flat tubing (LIT) sampler as a function of the product of aqueous diffusivity, log K0 and minimum molecular two-dimension. Another concern of using triolein-free polyethylene membrane as a passive sampling device is its relatively small sorption capacity due to lack of triolein (21, 22), subsequently the uptake phase would reach equilibrium during field deployment. In the present study, the linear relationship between the amounts of contaminants accumulated in LIT and SPMD and the results from PRC approach indicated both LET and SPMD were in the linear uptake phase. In addition, times for the phases of polyethylene membrane sampler uptake can be estimated from (16) tj = -in 0.5 K,w VPSD /R. (4) 91 where tj,2 is the time required to accumulate 50% of the equilibrium concentration which essentially represents the linear region of uptake, VPSD is the total volume of the PSD. Using LFT' sampling rates in Table 3.4, the average deployment durations for compounds with 4.4 log K 7.7 to maintain in the linear uptake phase during LFT deployment are 2,250 days at 9 °C (range from 29 to 9,810 days) and 1,280 days at 22 °C (range from 24 to 6,860 days. Evaluated together, our results confirm LFT sampler would not reach equilibrium during 21day exposure at realistic environmental concentrations even at a highly contaminated site with a wide array of contaminants, such as Portland Harbor superfund site. In summary, the triolein-free polyethylene lay flat tubing (LFT) showed the ability to detect the selected hydrophobic contaminants as efficient as the SPMD. Its sampling efficiency suggested the neutral lipid reservoir (triolein) present in SPMD was not always necessary. Also, biofouling was not a problem for its field application as compared to SPMD. This simpler device tended to accumulate compounds with high log IK faster than SPMD. Approaching equilibrium appeared not be a concern at realistic environmental concentrations for 21-day exposure. Spiking permeability/performance reference compounds can be achieved by using simple spiking method in a small volume of toluene. The successful determination of field derived data illustrated the effectiveness and reliability of the trioleinfree polyethylene lay flat tubing for environmental monitoring. Its application increases the range of tools available to those involved in monitoring water quality. The method of choice for environmental monitoring will depend on the contaminant nature and environmentally relevant concentrations of contaminants as well as the objective of a study. Acknowledgments. This study was partially funded by the Society of Environmental Toxicology and Chemistry (SETAC) Early Career for Applied Ecological Research Award sponsored by the American Chemistry Council to K.A.A. The Royal Thai government scholarship to D.S. is acknowledged. We wish to thank R. Grove for his field assistance. We would like to express appreciation for field and laboratory assistance by G. Sower, L. Quarles and J. Basile. 92 Literature Cited Huckins JN, Tubergen MW, Manuweera GK. Semipermeable membrane devices containing model lipid: a new approach to monitoring the bioavailability of lipophilic contaminants and estimate their bioconcentration potential. Chemosphere 20:533552(1990). Wells JB, Lanno RP. Passive sampling devices (PSDs) as biological surrogates for estimating the bioavailability of organic chemicals in soil. 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Chem. 14:18751884(1995). 95 Rantalainen A-L, Cretney WJ, Ikonomou MG. Uptake rates of semipeiiiieable membrane devices (SPMDs) for PCDDs, PCDFs, and PCBs in water and sediment. Chemosphere 40:147-158(2000). Schwarzenbach RP, Gschwend PM, Imboden DM. Environmental organic chemistry. New York, NY: John Wiley & Sons, Inc., 1993. Meadows JC, Echols KR, Huckins JN, Borsuk FA, Canine RF, Tillitt DE. Estimation of uptake rate constants for PCB congeners accumulated by semipermeable membrane devices and brown trout (Salmo trutta). Environ. Sci. Technol. 32:1847-1852(1998). Schwarzenbach RP, Gschwend PM, Imboden DM. Environmental organic chemistry. Hoboken, NJ: John Wiley & Sons, Inc., 2003. 96 CHAPTER 4 ENVIRONMENTAL STRESSES AND FISH DEFORMITIES IN THE WILLAMETTE RIVER: COMPARISON OF PERSISTENT BIOACCUMULATIVE TOXICANTS Doolalai Sethajintanin Eugene R. Johnson 1,2, and Kim A. Anderson Department of Environmental and Molecular Toxicology 2 Food Safety and Environmental Stewardship Program, Oregon State University, Corvallis, Oregon 97331 1,2 97 Abstract In response to public concern related to the reports of a high incidence of skeletal deformities in fish at the Willamette River, Oregon from River Mile 26.5-55, an area known as Newberg Pool, a team of multidisciplinary scientists from Oregon State University collaborated to identify the cause(s) of skeleton deformities associated with Willamette River fish, with particular emphasis on the Newberg region. Our research group's effort was to compare and contrast water chemical-physical conditions and the bioavailable fractions of PCBs, DDTs, and dieldrin at the Newberg and Corvallis areas, where fish deformities were significantly lower, and determine whether these factors were likely causes of the deformities. In situ monitoring of river water quality coupled with in situ sampling and analysis of bioavailable PCBs, DDTs, and dieldrin by semipermeable membrane device (SPMD) were used. There were some differences in water quality parameters between the Newberg and Corvallis sampling sites. However, they did not readily explain the difference in deformity loads. Concentrations of bioavailable PCBs, DDTs, and dieldrin were generally below 50 pg/L and the concentration differences observed between sites were very small, with the mean concentration differences ranged from none to 27 pgIL. The results did not provide compelling support for the hypothesis that PCBs, DDTs and dieldrin present in surface water or differences in water quality parameters were a likely cause of the different deformity loads observed at Newberg versus Corvallis. 98 Introduction In response to public concern related to the reports of a high incidence of skeletal deformities in fish at the Willamette River from River Mile 26.5-55 (an area known as Newberg) (1-3), a team of multidisciplinary scientists from Oregon State University collaborated to identify the cause(s) of skeleton deformities associated with Willamette River fish, with particular emphasis on the Newberg region (4). The incidence skeleton deformities in fish from the Newberg region had significantly greater deformity rates (56 %) than fish from the upper Willamette River (i.e., 2-5 % at Corvallis), see location in Figure 4.1 (2). Precaudal deformity load in northern pikeminnow showed a spatial pattern with a significantly higher incidence at Newberg area than the sampling sites at Newberg tributaries and the upper Willamette River(3). The high frequency of skeletal defects at the Newberg area and the gradual decline in the upstream and downstream direction suggested a local source as the cause of skeleton defects in this region. The collaborative study consisted of three main components as follows; investigation prevalence of skeleton deformities in juvenile fish from the Willamette River focusing on Newberg (high prevalence) and Corvallis (low prevalence) sites; comparison and contrast water physical and chemical conditions and distribution of selected chemical contaminants at these sites; laboratory toxicity experiments with complex chemical mixtures and other stressors from these sites. Together, these components provided a weight of evidence based empirical approach to identify the likely cause of skeletal deformities in Willamette fish. The final report has been published in Environmental Science and Technology. Volume 39, No. 10, 2005, page 3495-3506, "Environmental stresses and skeletal deformities infishfrom the Willamette River, Oregon" (4), see Appendix E. Our research group's effort focused on the distribution of persistent bioaccumulative toxicants in surface water. We compared water quality and potential for direct exposure to these chemical contaminants at the Newberg sampling site, north and south side of the area at River Mile 44 and 47, respectively, compared to the upper Willamette River sampling site at Corvallis (River Mile 135) and determined whether these factors were likely causes of the deformities. Inclusion of persistent bioaccumulative toxicants to be analyzed was based on in-depth review of the peer-review literatures. A wide variety of chemical, physical, and biological stressors have been associated with skeleton deformities in fish (5-10). This 99 chapter focuses on the distribution of chemical contaminants including polychlorinated biphenyls (PCBs), DDTs (l,1-bis(4-chlorophenyl)-2,2,2-trichloroethane and its metabolites) and dieldrin which have been reported to cause skeleton deformities by impairing developmental processes and bone formation (6-8). Smith and Cole (6) found embryo developing from eggs laid by adult winter flounder that were exposed to DDT showed a high incidence of vertebral deformities caused by bone erosion and hemorrhaging at the vertebral junctures. Olsson et al. (7) found offspring zebrafish following maternal exposure to certain PCB congeners showed craniofacial and posterior malformation. Skeletal deformities have also been linked to water quality problems, including low pH (5), low dissolved oxygen (11), and elevated temperature (12). The Willamette Bacin N D major tributary enter5 river Primary study sites City lucaltuire for eograptiic reference Direction of flow Figure 4.1 The Willamette Basin showing the general location and course of the Willamette River and primary study locations. The main purpose of this work was to compare and contrast water chemical-physical conditions and the bioavailable fractions of PCBs, DDTs, and dieldrin at the Newberg and Corvallis and determine whether these factors were likely causes of the deformities. In situ 100 monitoring of river water quality coupled with in situ sampling and analysis of bioavailable PCBs, DDTs, and dieldrin by semipermeable membrane device (SPMD) were used. SPMD consists of polyethylene lay flat tubing containing a thin film of a neutral lipid, 95% pure triolein (1,2,3-tri[cis-9-octadecenoyl]glycerol) (13). It has been proposed that SPMD mimics key mechanisms of bioconcentration including diffusing through biomembranes and partitioning between organism lipid and the surrounding medium (14). During spawning/early fish development (mid May through July 2002 and 2003), the in situ water quality monitoring device (YSI 6920 SONDE; Yellow Springs, OH) and SPMD were deployed to continuously collect a set of physical/chemical data. In this way episodic events and other pulses of contaminants were captured. Temperature, dissolved oxygen, pH, specific conductivity, oxidation reduction potential, NH-N,and NO3-N concentrations were collected on an hourly basis during each 21-day sampling event. Materials and Methods Passive sampling device. Standard SPMDs were purchased from Environmental Sampling Technologies (EST, St. Joseph, MO). A standard size SPMD consists of a 9 1-106 cm segment of 2.5 cm wide low density polyethylene lay flat tubing having a wall thickness of 70-95 tm and a surface area of 450 cm2, that contains 1 mL of 95% pure triolein as a thin film and has a total weight of 4.5 g (14, 15). Chemicals and solvents. Standards of organochlorine pesticides (purities 98.5%) were from Chem Service (West Chester, PA). Standard polychlorinated biphenyls (PCBs) (purities 99%) were from AccuStandard (New Haven, CT). All solvents used were pesticide or Optima® grade from Fisher Scientific (Fairlawn, NJ). Sample collection. The Willamette River sampling sites were chosen to facilitate investigation of spatial and seasonal bioavailable concentrations of organic contaminants at Newberg and Corvallis during May to July 2002 and 2003 (Figure 4.1.). Two sampling sites were located at Newberg; one on the south of the river (River Mile 47) and one a few miles downriver on the north side of the river (River Mile 44). Two sampling sites were located at 101 Corvallis at River Mile 135. SPMDs and YSI 6920 Sonde (YSI, Yellow Springs, OH) were deployed from May to July 2002 and 2003 to sample dissolved, bioavailable PCBs and organochiorine pesticides and to collect water chemistry parameters, respectively. Three 21day sampling events were completed per year, one each in May, June, and July. The sampling coincided with critical fish life stages during spawning and early development. River depth and water chemistry parameters including temperature, dissolved oxygen, pH, specific conductivity, oxidation reduction potential, NH-N,and NO3-N concentrations were measured on an hourly basis by YSI 6920 Sonde. Five individual SPMI)s were in a flow-through stainless steel cage. Each cage was suspended 1 ft. from the river bottom with submerged with "anchor-cable-cage-cable-float" arrangement. The five SPMDs were later composited for analysis. On retrieval, SPMDs were cleaned by gently rubbing with gloved hands in on-site water, then rinsed in iN HCI (approximately for 15 sec) (Trace metal grade, Fisher Chemical, Fairlawn, NJ), 18 MQ.cm water, acetone (Pesticide grade, Fisher Scientific, Fairlawn, NJ), and isopropanol (Pesticide grade, Fisher Scientific, Fairlawn, NJ), respectively. Cleaned SPMDs were kept in clean glass amber jars and transported on ice-packs. Samples were stored at -20°C until analysis. Sample extraction and chemical analysis. SPMD extraction and cleanup were described in detail in Chapter 2 and 3. Briefly, SPMDs were extracted by hexanes dialyses (Pesticide grade, Fisher Scientific, Fairlawn, NJ) in airtight amber glass jars. Sample volumes were reduced using rotary evaporation and TurboVap® LV evaporator (Zymark®, Hopkinton, MA). The samples were cleanup and fractionated using gel permeation chromatography (Waters® gel permeation chromatography cleanup system; Water® 515 HPLC pump, 717 Plus autosarnpler, Phenogel® column, 2487 dual X absorbance detector, and fraction collector II) (Milford, MA) with dichioromethane (Opima®, Fisher Scientific, Fairlawn, NJ) as the mobile phase at the flow rate of 5 mLlmin. Appropriate fractions were determined by analyzing standards and fortified samples. The fraction contained PCB and organochlorine pesticides was subjected to volume reduction using TurboVap® LV evaporator and solvent exchanged into iso-octane (Pesticide grade, Fisher Scientific, Fairlawn, NJ) and were analyzed using GC-pECD (Agilent Technologies 6890N Network 102 GC system) (Palo Alto, CA) dual capillary columns (db-xlb and db-17, J&W Scientific Inc., Agilent Technologies, Palo Alto, CA) /dual detectors. Quality control. Field duplicates and field blanks accompanying the deployed passive sampling devices during deployment, retrieval, and transportation to the laboratory were included in each sampling event. The Corvallis site was designated as a field duplicate site. These field blanks and laboratory control blanks represented 30 to 40% of a sample set and were processed and analyzed exactly as the deployed samples. None of the target analytes were identified in field blanks and laboratory control blanks. The percent recoveries of fortified samples were as follows: PCBs 57-110%, and organochiorine pesticides 56-72%, relative percent difference between field duplicates was 46%. PCB 170 and methoxychlor were used as permeability/performance reference compounds (PRCs), which were added to SPMD prior membrane enclosure (16). The use of PRCs as an independent measure for the exchange kinetics between SPMDs and water allows the device uptake rate adjustment for changes in temperature, flow velocity of the surrounding water, and biofouling on the membrane surface during field exposure (16, 17). The mean clearance rates of PCB 170 and methoxyclor indicated no significance loss during SPMD deployment. Biofouling or algae growth on SPMDs was nil to minimal in this study. No adjustment of sample concentrations for laboratory percent recoveries and percent loss in field was taken. Data analysis. The basic theory and mathematical models required for estimation of analyte water concentrations from the concentrations in the SPMD have been described by Huckins et al (15). Estimates of water concentrations (Cm) from SPMD concentrations were calculated by the following equation (15), C = CSPMD MSPMD R -1 where C SPMD is the concentration of the individual analyte in the SPMD, MSPMD is the mass of SPMD in grams, R is the sampling rate of a standard 1-g triolein SPMD, and t is the time 103 in days. SPMD sampling rates (R) for PCBs and organochiorine pesticides at multiple temperatures have been established (18-20). Data interpretation was performed using SPSS® Version 10.0.1(SPSS Inc., 19891999), Sigma Plot 2002 for Windows Version 8.0 (SPSS Inc., 1986-2001), and Microsoft® Office Excel 2003 (Microsoft Corporation, 1985-2003). Standard descriptive statistics and the Mann-Whitney test (nonparametric two-independent-samples tests) were used to evaluate differences between sampling sites and sampling year. Statistical analyses were considered significant atp 0.05. Results and Discussions Water chemistry characterization. Temperature, dissolved oxygen, pH, oxidation reduction potential, specific conductivity, NH4tN,and NO3-N concentrations were collected hourly during all 21-day sampling events in May-July, 2002 and 2003 (Table 4.1). Discrete sampling with 4-second interval was also measured during SPMD deployment and retrieval and was reported when the 21-day data were unavailable. At each sampling site, some parameters showed spatial and temporal variation while others did not. The temperature patterns at Newberg and Corvallis sampling sites were similar for both years and were generally 14 °C in May and rose to 20 °C by the end of the sampling period. The average river temperature at Newberg sampling site was generally 1-2 °C higher than at Corvallis. Diurnal temperature variation was similar at both sites (1-2 °C or less). Dissolved oxygen probe tended to drift after about 10 days of deployment in some sampling events. In that case the data after 10 days were excluded from the analysis. The dissolve oxygen pattern was similar at both sites. A slight decrease in dissolved oxygen was seen from May to July in both 2002 and 2003, as can be expected as water temperature increased. 104 Table 4.1 Spatial and temporal variations of water quality parameters at Newberg (River Mile 44-47) versus Corvallis (River Mile 135) sampling sites on the Willamette River, Oregon. water quality parameter temperature (°C) year 2002 month Newberg Corvallis May 14 ± 1.2 13 ± 0.93 June 16±1.3 16±1.3 July 19 ± 1.3 19 ± 1.5 15±0.Oa 2003 May 14± 1.7 June 17±1.1 16±1.3 July 20±2.0 19± 1.9 dissolved oxygen (mgJL) 2002 May 12 ± 0.49 11 ± 0.52 June 9.9 ± 0.61 9.2 ± 1.4 July 9.6 ± 0.35 9.5 ± 0.39 12±0.28a 2003 May 1o±o.o June 11± 0.8 10±0.41 July 9.7±0.41 9.6±1.6 pH 2002 May 7.5 ± 0.06 7.6 ± 0.16 June 7.5±0.12 7.6±0.26 7.4 ± 0.01 July 7.3 ± 0.04 2003 7.4±0.0 7.4±0.12 May June 7.4±0.16 7.7±0.4 7.4±0.1 July 7.2±0.10 260 ± 26b 370 ± 16b oxidation-reduction 2002 May 120 ± 13b 340 26b potential or ORE' (mV) June 29017b 44033b July 27040b 165 ±2.3 a 2003 May 38087b 29032b June 44020b 39036b July specific conductivity 2002 May 0.063 ± 0.00 1 0.060 ± 0.002 (mS/cm) June 0.057 ± 0.004 0.067 ± 0.004 July 0.078 ± 0.004 0.075 ± 0.003 2003 0.073 ± 0.002 0.064 ± 0.000 a May June 0.060 ± 0.006 0.063 ± 0.004 0.06 1 ± 0.002 July 0.087 ± 0.002 0.05 ± 0.01 2002 0.04 ± 0.01 May NH-N (mg/L) 0.04 ± 0.01 0.09 ± 0.01 June July 0.17±0.03 0.16±0.01 0.10±0.Oa 2003 0.04±0.01 May June 0.10±0.02 0.07±0.02 July 0.03 ± 0.01 0.02 ± 0.00 2002 May NA NA NO3-N (mgIL) NA June NA July 0.86 ± 0.05 a 1.17 ± 0.16 a 2003 May 1.1 ± or' 2.8 ± 0.30 a June NA NA July 0.36 ± 0.14 a 0.2 ± 0.0 a a Continuous sampling data were not available. Discrete sampling data taken during SPMD deployment and retrieval were reported. b0 probe tended to drift after 7-10 days of deployment. Data after 7 data were excluded from the analysis. NA- data not available 105 Maximum pH range, usually in late May to mid June was 7.4 to 8.6 at Corvallis sampling site, and 7.2 to 7.8 at Newberg sampling site. The daytime pH showed a strong spatial difference, with the Corvallis sampling site often 1 + pH units higher than the Newberg sampling site. By contrast, the night time pHs were similar at both sites. These large diurnal pH changes were seen in both years. There were small decreases in pH from 2002 to 2003. Although some deformities in fish have been linked to pH effects such as muscular and spinal deformities due to decalcification in white sucker held at pH 4.2 for over 4 weeks (5), we are unaware of any peer-reviewed literatures linking pH 7.2 to 8.6 conditions to skeleton deformities in fish. The oxidation reduction potential (ORP) probe tended to drift after 7-10 days of deployment. Since all deployments were 21 days, data after 7 days were excluded from the analysis. In 2002, the ORP was lower at the Newberg sampling sites as compared to the Corvallis sampling site with the difference ranged from < 10% to a factor of 2. In contrast, the ORP at Newberg was higher than Corvallis in 2003. There was some evidence that ORP differed among years at the Newberg sampling sites. The ORP value is a direct reading of the activity of oxidizing and reducing agents in the water as they correspond to oxidationreduction reactions. In general, there was no specific ORP pattern difference between Newberg and Corvallis sampling sites although the Newberg waters were observed more reducing than Corvallis waters in 2002. ORP is known to influence microbial growth, but it was not clear whether the difference in ORP observed would make Newberg fish more vulnerable to infection by microbial agents or more susceptible to deformities. Specific conductivity was similar at the Newberg and Corvallis sampling sites for both years. A seasonal increase in NH4tN concentrations was observed at both sites from May to early July, with Corvallis generally having higher NH4-N than Newberg. The NO3N probe was not robust. In 2002, drift generally occurred within 24 hours of deployment. In 2003, the probe failed within a few hours of deployment. The limited data indicated that NO3 -N decreased from May to July, the concentrations were higher at the Newberg sampling sites as compared to the Corvallis sampling sites. Evaluated together, water chemistry parameters at both sites provided no compelling evidence to suggest that the differences in 106 these parameters were likely causes of the different fish deformity loads observed at Newberg versus Corvallis. Bioavailable polychiorinated biphenyls (PCBs). In the present study, PCB analysis was based on a congener-specific approach; however, concentrations of all congeners were quite low and were below detection limits (0.71-4.4 pgIL depending on congeners) in many samples, so interpretation of results focused on PCB concentrations (sum of 24 congeners). The bioavailable concentrations of PCBs were generally very low at all sites (Table 4.2). During 2002 and 2003, biovailable PCB were generally greater at Newberg sampling sites (Table 4.2). However, since most of the data were near or below detection limits, it was difficult to draw any firm conclusions. Bioavailable concentrations of PCBs were approximately 500-fold lower than the national and the Oregon fresh water aquatic life criteria for PCBs (0.0 14 tgIL) (2], 22) and they did not readily explain the difference in deformity load among sites. Table 4.2 Mean concentrations (pg/L) of bioavailable organochlorines estimated from concentrations accumulated in semipermeable membrane devices (SPMDs) exposed for 21 days at the Willamette River at Newberg and Corvallis (2 sampling sites per location) in 2002 and 2003 temporal spatial distribution chemical 2002 NP CV NP CV n=6 n=6 p-value p-value NP CV 8.7±4.0 0.015 1.000 0.394 48 ± 11 36 ± 4.2 0.065 0.485 0.240 0.041 11±2.2 11±0.99 0.394 1.000 0.004 6.2±7.6 0.041 21±5.9 16±2.7 0.132 0.180 0.041 13 ± 19 0.18 15 ± 3.9 9.7 ± 1.6 0.026 0.180 1.000 8.1 ± 9.0 0.015 25 ± 5.9 14 ± 1.8 0.004 0.394 0.818 n=6 PCBsd 13±15 7.0±11 0.39 15±5.2 DDTs 49 ± 18 22 ± 28 0.18 p,pDDTc 10±5.2 p,pDDEc 16±5.3 p. pDDDd 23 ± 8.7 2.9±4.6 23 ± 6.8 p-value 2003 n=6 dieldrin" distribution asee chapter 1 for complete list of analyte. b significant difference between sites, both years. significant difference between sites, 2002 only. significant difference between sites, 2003 only. 107 p, p'-DDT and its metabolites. The bioavailable DDT (sum of p, p'-DDT, p, p'- DDD and p. p'-DDE) concentrations were vary low at all sites in 2002 and 2003 (Table 4.2). Bioavailable DDT concentrations were approximately 10-fold below the national and the Oregon fresh water aquatic life criteria 0.001 pgIL (21, 22) which is the level generally thought toxic to fish. The differences in bioavailable DDT between Newberg and Corvallis sites in 2002 (p=O. 18) and in 2003 (p=O.O65) were not significant. In 2002, the bioavailable DDT profile was dominated by p, p'-DDD, followed by p. p'-DDE and p. p'-DDT (Figure 4.2). In 2003, bioavailable p, p'-DDE dominated, followed by p, p'-DDD and p, p'-DDT. This would be expected from older deposits and environmental degradation of p, p'-DDT to its metabolites. Bioavailable p. p'-DDD and p. p'-DDE concentrations detected at Newberg sampling sites in 2002 and 2003 were not significantly different (p=O.l8) (Table 4.1). Bioavailable p. p'-DDE concentrations but not bioavailable p. p'-DDD concentrations at Corvallis sampling sites increased significantly between 2002 and 2003 (p=0.04l for p, p'- DDE and p=l for p, p'-DDD). The mean concentrations of bioavailable p. p'-DDT at Newberg did not increase significantly between 2002 and 2003 (p=l), but the mean concentrations of bioavailable p. p'-DDT detected at Corvallis increased significantly (p=O.004). The cause of the increased p. p'-DDT concentration was not clear, but the results suggest possible new inputs or sediment disruption remohilizing DDT. However, the concentration differences observed were very small, so conclusions should be drawn prudently. 80 -J 60 C Newberg Newberg 0 1 Corvallis 0 40C 0 0 p. p'-DDE p. p'-DDD p. p-DDT Corvallis C 0 0 2002 2003 Figure 4.2 The mean concentration distribution of bioavailable p, p'-DDT, p. p'-DDD and p. p'-DDT at Newberg and Corvallis in 2002 and 2003 108 Dieldrin. Bioavailable dieldrin concentrations were significantly higher at Newberg sampling sites as compared to Corvallis sampling sites (p=O.OlS for 2002 and p=0.004 for 2003) (Table 4.1). However, the bioavailable concentrations of dieldrin were very low and there was no significant difference between years. The bioavailable dieldrin concentrations detected were approximately 2000-fold below the national and the Oregon fresh water aquatic life criteria, 0,056 tgIL and were well below those generally thought to be toxic to fish (23). We are not aware of any reports linking picogram-per-liter concentrations of dieldrin to skeletal deformities. To draw causation of fish deformities at Newberg due to chemical stresses, we can eliminate compounds that are absent, or if present, their concentrations do not differ from the Corvallis sampling sites where fish deformities are relatively low. The results indicated the difference between the presence or absence of bioavailable PCB congeners, p,p' - DDT and its metabolites, and dieldrin detected at the Newberg and Corvallis sampling sites was insignificant. For the compounds that were present there appeared to be little difference in their water concentrations between the sites. The results did not provide compelling support for the hypothesis that PCBs, DDTs and dieldrin present in surface water were a likely cause of the different deformity loads observed at Newberg versus Corvallis. The results coincided with the concentrations of these compounds found in fish ovarianloocyte and sediment samples (4). Most organochiorine compounds were below detection limits in fish ovarianloocyte and sediment samples and those that were detected were not significantly different between sites. In contrast, the results from the other collaborative group found metacercariae of a digenean trematode were directly associated with a large percentage of deformities detected in Willamette River fish and similar deformities were reproduced in laboratory fathead minnows exposed to cercariae extracted from Willamette River snail, see detail in Appendix E. Therefore, the weight of evidence suggested that parasitic infection, not chemical contaminants, was the primary cause of skeleton deformities observed in Willamette River fish. Acknowledgments. This part of the project was supported by a grant from the Oregon Watershed Enhancement Board (no. 20 1-562). The Royal Thai government scholarship to 109 D.S. is acknowledged. We wish to thank R. Grove for his field assistance. We would like to express appreciation for field assistance of J. Basile, O.Krissanakriangkrai and R. Adams. Literatures Cited Ellis SG. Characterization of skeletal deformities in three species of juvenile fish from the Willamette River basin EVS Project No. 2/839-02. Seattle, Washington: Report for the Oregon Department of Environmental Quality (ODEQ) by EVS Environmental Consultants, Inc., 2000. Ellis SG, Deshler ST, Miller R. Characterizing fish assemblages in the Willamette River, Oregon, using three different bioassessment techniques. In: River quality dynamics and restoration (Laenen A, Dunnette DA, eds). New York, NY: Lewis Publishers, 1997;347366. Markie DF, French R, Naman S, Demarest J, Hatfield M. Historical patterns of skeletal deformities in fishes from the Willamette River, Oregon. Northwestern Naturalist 83:714(2002). Villeneuve DL, Curtis LR, Jenkins JJ, Warner KE, Tilton F, Kent ML, Watral VG, Cunningham ME, Markie DF, Sethajintanin D, Krissanakriangkrai, Johnson ER, Grove RA, Anderson KA. Environmental stresses and skeletal deformities in fish from the Willamette River, Oregon. Environ. Sci. Technol. 39:3495-3506(2005). Beamish RJ. Lethal pH for the white sucker (Catostomus commersoni) (Sapede). Trans. Am. Fish. Soc. 101:355-358(1972). Smith RM, Cole CF. Effects of egg concentrations of DDT and dieldrin on development in winter flounder (Pseudopleuronectes americanus). J. Fish Res. Board Can. 30:18941898(1973). Olsson P-E, Westerlund L, Teh SJ, Bilisson K, Berg AH, Tysklind M, Nilsson J, Eriksson L-O, Hinton DE. Effects of maternal exposure to estrogen and PCB on different life stages of zebrafish (Danio rerio). Ambio 28: 100-106(1999). Mehrle PM, Haines TA, Hamilton S, Ludke JL, Mayer FL, Ribick MA. Relationship between body contaminants and bone development in east-coast stripped bass. Trans. Am. Fish. Soc 111:231-241(1982). Kent ML, GroffJM, Morrison JK, Yasutake WT, Holt RA. Spiral swimming behavior due to cranial and verterbral lesions associated with Cytophaga psychrophila infections in salmonid fishes. Dis. Aquat. Org. 15:115-121(1989). Kent ML, Watral V. Whipps C, Cunningham ME, Criscione CD, R HJ, Curtis LR, Spitsbergen J, Markie DF. A digenean metacercariae (Apophallus sp.) and a myxozoan (Myxobolus sp.) associated with vertebral deformities in cyprinid fishes from the Willamette River, Oregon. J. Aquat. Anim. Health. 16:116-129(2004). 110 Blaxter JHS. Development: eggs and larvae. In: Fish physiology, Vol 3 (Hoar WS, Randall DJ, eds). New York, NY: Academic Press, 1969;177. Brungs WA. Chronic effects of constant elevated temperature on the fathead minnow (Pimephalespromelas Rafinesque.). Trans. Am. Fish. Soc 4:659-664(1971). Huckins JN, Tubergen MW, Manuweera GK. Semipermeable membrane devices containing model lipid: a new approach to monitoring the bioavailability of lipophilic contaminants and estimate their bioconcentration potential. Chemosphere 20:533552(1990). Petty JD, Orazjo CE, Huckins JN, Gale RW, Lebo JA, Meadows JC, Echols KR, Cranor WL. Considerations involved with the use of semipermeable membrane devices for monitoring environmental contaminants. J. Chromatogr. A 879:83-95(2000). Huckins JN, Petty JD, Lebo JA, Orazio CE, Clark RC, Gibson VL. SPMD technology tutorial (3' Edition). Last update January 3, 2002 {cited July 5, 2004]. Available from http://wwwaux .cerc.cr. usgs .gov/spmd/SPMD-Tech Tutorial .htm Huckins JN, Petty JD, Lebo JA, Almeida FV, Booij K, Alvarez DA, Cranor WL, Clark RC, Mogensen BB. Development of the permeability/performance reference compound approach for in situ calibration of semipermeable membrane devices. Environ. Sci. Technol. 36:85-91(2002). Booij K, Sleiderink HM, Smedes F. Calibrating the uptake kinetics of semipermeable membrane devices using exposure standards. Environ. Toxicol. Chem. 17:12361245(1998). Meadows JC, Echols KR, Huckins JN, Borsuk FA, Canine RF, Tillitt DE. Estimation of uptake rate constants for PCB congeners accumulated by semipermeable membrane devices and brown trout (Salmo trutta). Environ. Sci. Technol. 32:1847-1852(1998). Rantalainen A-L, Cretney WJ, Ikonomou MG. Uptake rates of semipermeable membrane devices (SPMDs) for PCDDs, PCDFs, and PCBs in water and sediment. Chemosphere 40:147-158(2000). Petty JD, Huckins JN, Orazio CE, Lebo JA, Clark RC, Gibson VL. Evaluation of the semipermeable membrane devices (SPMD) as a passive in situ concentrator of military organic chemicals in water, Final report, approved for release. Fort Dentrick, MD: US Army Medical Research and Material Command, 1997. The United States Environmental Protection Agency. National recommended water quality criteria: 2002 EPA-822-R-02-047. Washington, DC: Office of Water and Office of Science and Technology, 2002. Fitzpatrick MS. Toxic compounds criteria: 1999-2003 Water quality standards review, issue paper. EQC Meeting, Agenda item B, Rule adoption. Portland, OR: The State of Oregon Department of Environmental Quality, 2004. WHOIIPCS. Aidrin and dieldrin. Geneva:World Health Organization, 1989. 111 CHAPTER 5 CONCLUSIONS As part of an assessment of the bioavailability of a chemical contaminant, this dissertation demonstrated the seasonal distribution of bioavailable organochiorine contaminants in a river system and evaluated the potential environmental factors influencing their bioavailability. The study was carried at the lower Willamette River at Portland Harbor, Oregon where surface water runoff is determined by climate influence and varies according to season. Bioavailable water concentrations of PCBs, DDTs, and dieldrin were determined using polyethylene membrane containing passive sampling device, known as semipermeable membrane device (SPMD). Our findings indicated that the influence of river seasonality on the bioavailable distributions of organochlorine contaminants was compound- and sitespecific. Seasonal variation of bioavailable DDTs was site-specific. A seasonally significant increase of bioavailable DDT concentrations was observed during low flow in summer at the sampling site just downstream of the former DDT production and handling facilities. The large bioavailable concentrations of DDTs coincided with large DDDIDDE ratios. The ratio of DDDIDDE was highly variable, 6.6 ± 2.1 in summer, and 2.8 ± 1.3 in fall. A significant increase of bioavailable p. p'-DDD during low flow in summer was a major driver of seasonality in bioavailable DDT concentrations. The linear regression analyses suggested seasonal changes in bioavailable p, p'-DDD concentrations were potentially related to changes in water temperature, river flow, and precipitation. Slow moving surface water, declining redox condition, elevated water temperature and decreased oxygen concentration during summer as observed in the present study would enhance anaerobic reductive dechlorination of p, p'-DDT to p, p'-DDD by both microbial degradation and chemical reaction in the sediment. In addition, the effect of increasing temperature in summer could decrease the organic matter-water partition coefficient of contaminants, resulting in 112 increasing bioavailable DDTs release from the contaminated sediment. Evaluated together, increasing formation and release of p, p' -DDD in sediment during low flow in summer would contribute to the large increase of bioavailable p, p'-DDD and large ratio of DDD/DDE in surface water. As water temperature decreased and river flow increased in fall, the condition suitable for reductive dechlorination may have been altered, resulting in decreased bioavailable p, p'-DDD concentrations and change in bioavailable DDDIDDE ratios. The above discussion contains speculation of the processes that highlights the lack of understanding on how seasonal river conditions interact to yield the observed seasonal variation in bioavailable p, p'-DDD and its relationship to bioavailable p, p'-DDE. Further research is needed such as confirmation of microbial communities responsible for reductive dechlorination of p, p ' -DDT to p, p ' -DDD or reductive sediment condition and their role in seasonal bioavailable p, p'-DDD distribution. Understanding the interaction of seasonal physico-chemical and biological factors will afford confidence in using this knowledge to predict the long-term seasonal effects of DDTs on aquatic community and increased confidence in extrapolating information to other contaminated sites. Bioavailable PCB concentrations were found to have strong spatial and seasonal variation. Elevated concentrations of bioavailable PCBs usually reached a maximum at the sampling sites within the industrial area. Both concentrations and daily loads of bioavailable PCBs were significantly increased during high flow in fall, especially episodic rainstorms and the discernable seasonal pattern was observed at almost every sampling site along the 18mile stretch of the study area. The results suggested the sources of bioavailable PCBs in surface water at the lower Willamette River were presumably seasonally dependent. Storm flow was especially important. Our results suggested storm events may have disturbed the interaction between PCBs and their associated organic matter resulting in remobilization of bioavailable fraction from sediments or particulate matter. In addition, the increase in bioavailable PCB concentrations and daily loads which was coincident with high precipitation and sewer overflows in fall suggested a significant contribution of PCBs from precipitation input, urban storm water and urban community wastewater discharges to the surface water. 113 The hypothesis that combined sewer overflows and precipitation could be a major source of PCBs during high precipitation and high river flow in the lower Willamette River warrants further investigation such as PCB fingerprinting to fully elucidate their contribution to PCB contamination. Further studies will be necessary to estimate the impact of the internal sources (i.e., contaminated sediments) and other sources (i.e., precipitation, runoffs, wastewater discharges) of PCBs and how much they contribute to the PCB problem in the system in order to reduce the PCB load and risk to the aquatic organism community. Although no discernable spatial pattern was observed for dieldrin, an increase in bioavailable dieldrin concentrations and daily loads was observed at most sampling sites during high flow/high precipitation in fall. The seasonal pattern of bioavailable dieldrin distribution illustrated the transport behavior for non-point source contaminants which were widely dispersed and had no intentional seasonal application cycle such as with the current use pesticides. Our results combined with the historical use of dieldrin in the watershed suggested bioavailable dieldrin transport from contaminated site upstream including erosional inputs to the river during precipitation runoff from land sources or riverbed sediment. To screen the potential impacts associated with seasonal changes of bioavailable organochlorines on aquatic organisms, the bioavailable concentrations of DDTs, PCBs and dieldrin were compared to the national recommended water quality criteria and the Oregon water quality criteria. Seasonal exceedances of the criteria for DDTs were limited to only the sampling site downstream of the former DDT production and handling facilities during low flow in summer. Exceedances of the criteria for dieldrin were frequently observed during high flow in fall. In contrast, no discernable seasonal pattern of criteria exceedance was observed for PCBs. These findings suggested the potential impacts associated with seasonal changes of bioavailable organochiorine distributions in surface waters and the significances of considering seasonal, chemical-, and site-specific conditions in risk assessment and water quality management. 114 Importantly, predicting bioconcentration from measured water concentrations of bioavailable organic contaminants using passive sampling devices can provide more accurate bioconcentration factor (BCF) compared to using conventional filtered water methodology. Measured or predicted BCFs are an essential component of both human and environmental risk assessment because BCFs illustrate the importance linkage between the water column and its influence on exposure of aquatic organisms. BCFs are strongly influence prediction of toxic effects, especially when chemical residue-based dose-response relationships are used (Barron, 1990 and Cook and Burkhard, 1996). Many hydrophobic organic contaminants such as PCBs and organochiorine pesticides have a high affinity for particulates and dissolved organic matter, which can reduce bioavailability of chemicals, Predicting BCFs from dissolved water concentrations using conventional filtered water methodology would include contaminants associated with dissolved organic matter, which could not transport across biomembranes. Thus, it will underestimate the BCF. In contrast, predicting BCFs using passive sampling devices (BCFPSD) accounts for the bioavailability consideration and consequently reduce uncertainty for site-specific bioavailability conditions. The present study assesses the sampling device attributes. For instance, the passive sampling technique can be successfully used to assess spatial and seasonal distribution of bioavailable organochiorines in surface water, as well as, to allow a direct assessment of the water quality based on bioavailable concentrations, as presented in Chapter 2. Also, it can be used as a tool to assess the potential for direct exposure to hydrophobic organic contaminants and to determine whether these contaminants are likely cause(s) of the adverse effect(s) observed in aquatic organisms, as presented in Chapter 4 and Appendix E. However, an inexpensive and simpler time-integrative passive sampling device is still needed. In response to a growing need of an alternative passive sampling device, our research group has further developed a triolein-free low density polyethylene membrane lay flat tubing (LFT). Our laboratory and field verification demonstrated that the LFT without triolein proved reliable and had the same benefits as SPMD, but was simpler, inexpensive and lacked the interference from triolein impurities as found in SPMD. Our results found that LFT tended to accumulate compounds with high log K0 faster than SPMD. 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Deployments of SPMDs were typically 7- to 21-day long. 127 p,p'-DDE p,p'-DDD p,p'-DDT (pg/L) (pg/L) (pgIL) 2-15 3-7 3-7 66 (5.3) 44 167 (49) 2.3 (3.3) 236 (51) 126 174 55 NA 250 514 NA 4 5 RM8 48 112 RM 12 15 18 11 13 59 56 20 46 11(4.1) 120 (1.5) 66 68 sampling event detection limit (pg/L) a August 2001 RM 1 RM3.5 West RM 3.5 East RM7 West RM 7 East RM13 RM 15 RM 17 RM 18c September 2001 RIvI 1 RM 3.5 West RM 3.5 East RM7 West kM 7 East 125 48 NA bdl bdl DDTs (pg!L) 309 687 NA 161 dieldrin (pgIL) PCBs (pg/L) 3-19 06954b 43 (2.0) 47 58 57 NA 75 42 13 (11) 14 31 31 NA 13 14 18 5 74 79 bdl 31 72 23 37 4.8 13 42(25) 7.5 (11) 60(40) 35 (15) 10(8.2) 289 (32) 327 6.0 (8.5) bdl bdl 414 (39) 393 439 65 (2.5) 50 120 (13) 94 100 NA 1257 55 108 NA 595 NA NA 71 123 12 13 11 134 115 184 41 8.5 89 201 371 956 NA NA 114 bdl bdl 33 481 112 73 144 61 18 111 34 50 38 63 RM 18c October 2001 21(0.77) 57 (68) 4.6 (6.5) 82 (60) 45 (3.0) 13 (4.8) RIvI 1 82 (10) NA NA 125 (18) NA NA 266 NA 35 25 34 32 (3.7) NA NA 239 (25) NA NA 56 (2.7) NA NA 59 NA 34 40 40 130 (0.35) NA NA 72 NA 36 29 26 13 20 28 (2.8) RM8 RM12 RM13 RM15 RM17 kM 3.5 West RM 3.5 East 22 28 30 RM7West 87 RM 7 East NA RM8 RM12 33 RM 13 27 23 RM15 RM17 RIvI 18c November 2001 RM 1 21 18 26(0.32) 71 73 kM 3.5 West NA NA RM3.5East RM7 West 114 93 RM 7 East NA 50 46 140 180 NA RM8 RM 12 RM 13 RM 15 RM 17 RM 18 July 2002 RM 1 R1v13.5 West 33 14 4.2 17 168 521 NA NA 14 83 18 10 64 79 45 31 16 13 51 26 21 16(0.12) 69 (2.4) 62(0.44) 35 (1.1) 170 75 NA 18 26 NA 76 76 NA 22 32 26 NA 27 42 29 30 NA 44(11) 95 (6.3) 7.1 (0.72) 54 159 13 67 56 50 63 21 18 NA NA 330 349 NA 104 105 135 106 98 NA 146 (18) 226 104 NA NA 270 94 NA 78 101 83 102 104 49 100 37 105 NA NA 24(1.2) 11(1.7) 54 33 137 81 101 128 sampling event detection limit (pg/L) a p,p'-DDE p,p'-DDD p,p'-DDT (pgIL) (pg/L) (pg/L) 2-15 3-7 3-7 147 14 143 212 1242 53 NA NA 13 126 76 NA 65 RM3.5East RM7 West 52 160 RM 7 East NA RM8 RM12 RM13 43 25 45 24 29 RM 15 RM 17 RM 18c August 2002 RIvI 1 RM3.SWest RM3.5East RM7 West RM7 East RM8 RM12 RM 13 RM 15 RM17 RM 18c September 2002 RM 1 RM 3.5 West RM3.SEast RM 7 West RM 7 East d RM8 RM12 RM13 RM15 RM17 RM18C November 2002 RM 1 939 NA 70 42 9 DDTs (pg/L) dieldrin (pgIL) PCBs (pg/L) 3-19 06954b 71 52 47 NA 35 51 13 74 38 50 44 (15) 32 6.4 8.5 68 8 30 31 8 7 121 63 68 18 (4.1) 24 (6.5) 4.3 (1.3) 47 (12) 52 (2.5)c 17 (1.8)c 154 (34)C 300 319 1465 28 (0.97)c 62 66 129 85 (2.6)c 218 229 1207 NA NA NA NA 37 23 25 23 23 18 (4.1) 63 33 12 112 10 41 33 31 11 66 76 48 41 47 13 65 67 46 60 25 (5.3) 10(2.4) 53 (12) 41(7.3) 12 (2.6) 49(2.5) 96(18) NA 175 18 759 68 (21) 74 20 (12) 156 (22) NA 245 923 126 (39) 74 26(2.2) NA 11(1.8) NA 32(5.6) NA 52 90 37 (10) 27 22 29 NA 56 52 55 (42) 24 61 66 20 24 130 NA 9 67 8.1 (2.7) 15 45 60 52 NA 37 19 26 9.3 11 11 58 78 43 39 27 (10) 28 28 30 7 49 23 8 39 22 3.0 bdl 22(1.3) 9.8(1.1) 50(1.7) 33(0.14) 9.5(1.6) 204 (16) 200 83 (8.0) 29(3.5) 69 65 32 37 30 16 (3.7) 39 7 11 21 25 38 22 15 16 18(0.61) 91(3.6) 102(11) 11(1.2) RM3.5 West 78 108 RM 3.5 East 79 RM7 West 61 129 14 14 321 RM 7 East C 43 (4.8) 41(5.4) 37 38 RM 15 45 47 RM17 153 55 7.6 (0.079) 505 92 (10) 28 24 7 9 13 35 27 7 9 6 73 70 88 83 27 13 5.6 29 4.9 bdl 52(8.0) 37(9.2) 10(1.1) 99(18) 84(24) 51(7.3) 76 (12) 14 (1.5) 36 37 260 (18) 332 47 (2.6) 54 RM3.SEast RM7West 170 (4.5) 242 242 426 48 (2.6) 54 59 105 338 61 149 61 47 (8.9) 51(12) 8.9 (6.9) 680 107 (25) 48 RM 7 East e 40 (22) 41(14) RM8 RM12 RM13 RM18C October 2003 RM 1' RM 3.5 West 8 64 50 (0.29) 46 46 53 53 12 22 10 69 102 129 sampling event detection limit (pg/L) a RM8 RM12 RM13 RM 15.5 RM 17 RIvI 18.5 November 2003 RM 1 RM 3.5 West RM 3.5 East RM7 West RM 7 East C RM8 RM 12 RM13 RM15 RM 17 RM18 July 2004 RM ic p,p'-DDE p,p'-DDD p,p'-DDT (pgIL) (pgIL) (pg/L) 2-15 3-7 3-7 24 NA NA 23 34 6 65 NA NA 26 NA NA 49 30 29 14 (0.53) 527 (35) NA 322 533 131 (13) 13 16 127 101 20 139 118 93 91 68 17 16 16 35 (2.7) 55 47 52 39 45 50 13 14 17 15 NA NA bdl bdl bdl 264 (17) NA 223 (8.1) NA 39 (9.6) NA 119 37 102 166 199 67 (6.7) 70 58 81 69 53 54 232 50 (6.0) 44 26 38 32 24 21 DDTs (pg/L) 102 (7.4) 234 9.0 (0.18) 161 (9.5) 21 RM3.5East 50 (1.9) 57 42 133 RM7 West 111 RM7East RM8 29 804 66 54 14 132 8 8 RM 12 28 RM13 RM15 RM17 28 23 RM3.5 West RM 18c August 2004 RM 1 RM 3.5 West RM3.5East RM7 West RM 7 East RM8 RM12 RM13 dieldrin PCBs (pgIL) (pg!L) 3-19 0.69- 54b 34 NA NA NA NA 35 24 29 bdl bdl 76 (5.9) NA 86 76 67 (2.8) 64 60 74 58 62 35 1.5 39 (3.5) NA 80 22 15 (6.7) 13 bdl 5.4 4.3 bdl 5.5 90 (6.3) 39 39 bdl 34 28 43 6 9 311 188 1046 103 88 67 75 58 86 19 (4.4) 22(2.9) 6.0 (1.3) 47 (8.5) 38 (7.6) 42 (3.3) 91(0.52) 126 (2.1) 56 (1.1) 95 (2.9) 149 120 25 273 (2.7) 236 68 (2.8) 63 52 106 NA 30 25 62 65 69 152 123 116 NA 67 68 60 68 NA 26 546 NA 50 7 29 161 201 813 NA NA 11 13 91 45 43 64 128 154 126 99 90 89 90 42 34 RM17 31 19 20 8 74 83 90 48 51 104 69 92 51 35 RM 18C 23 (0.23) 29(1.0) 15 (5.2) 67(4.0) 60(2.0) 53 (4.6) 122 (1.1) 70 139 (3.0) 108 (1.5) 260 (3.1) RM 3.5 East 82 28 (0.27) 29 29 290 (4.4) RM3.5 West RM7 West 282 71 93 105 RM7East RM8 31 152 408 184 63 95 64 RM 15 October 2004 RM 1 RM12 RM13 28 45 32 56 36 37 43 104 98 18 17 656 30 403 41 10 10 203 209 1341 69 96 71 21 129 28 53 8 66 85 69 84 162 130 sampling event detection limit (pg/L) a RM15 RM17 p,p'-DDE p,p'-DDD p,p'-DDT (pg/L) (pg/L) (pg/L) 2-15 3-7 3-7 49 39 10 37 27 47 (0.93) 9 18 (1.2) DDTs (pg/L) dieldrin (pg/L) (pg/L) 3-19 0.69-54' PCBs 46 49 RM 18c 58 (0.58) 124 (2.7) 68 (3.2) n = 1 composite sample of 5 SPMDs; duplicate samples were at River Mile 1 and 18; values in parenthesis are 1SD; bdl- below detection limit; NA-data not available a range of method detection limits from 13 sampling events b detection limits for 25 individual PCB congeners Cn d 2 19 15 98 73 86 86 123 (0.90) 131 Appendix B. Sewer overflow points in Portland area (City of Portland, 2004, http://www.portlandonli ne.corn/bes/index.cfm9c=29323) area of former POB using facilities RM COLUMBIA RIVER sewer overflow points RM sewer overflow points (99% eliminated on the Slough) / RM7 sampling sites COLUMBIA SLOUGH RM\ WILLAMETTE RIVER 4 RM13 RM 17 RM isL\ COMWNED SEWER AREA 132 Appendix C. Equations used to estimate molecular diffusitivity in water (D1 kT = Stokes-Einstein relation 67trr1 ) where k = the Boltzmann constant (1.381 x 1023 kg m2 s2 K1) = dynamic viscosity (kg rn1 T = temperature (K) r, = molecular radius = [3Vj/41tNA]L3 where V = molar volume of the chemical (cm3 moi') NA = Avogadro's number (6.02 x 1023 mol') = Hayduk and Laudie 13.26 x 10 1.14 v where = solution viscosity (102 g cm1 s') at the temperature of interest V = molar volume of the chemical (cm3 mol') 133 Appendix D. Predicted values for LFT sampling rates using different methods. 3.4 . 3.2 C, ci) C) predicted R from water diffusion coefficient by Strokes- Einstein at 9 °C -c :3 3.0 predicted R from water diffusion coefficient by Hayduk & Laudie at 9 °C . 4.8 4.4 4.0 S 2.8 ci) 0 ci) c. 5.2 S 3.6 ci) 2.6 S .. 3.2 2.4 2.8 100 150 200 250 300 350 100 150 molar volume (cm3/mol) 5.0 4.8 4.4 (:1 ci) Strokes-Einstein at 22 °C S 7.5 I C)) S -c S 4.2 - ci) 0 -c 4.0 ci) S S 3.8 - . ci S 3.6 100 150 200 250 300 . 300 350 7.0 predicted R from water diffusion oefficient by 6.5 Hayduk&Laudieat22°C 6.0 S 5.5 5.0 4.5 4.0 350 100 200 150 molar volume (cm3/mol) 250 300 350 molar volume (cm3!mol) 25 12 estimated RSLFT from R$sPMD 10. -j 250 200 molar volume (cm3/mol) predicted R from water diffusion coefficient by S 4.6 - ci) S. S estimated RSLFT from RSsPMD -c :3 S 8- U- I S ci) 20- S $ 15S . 2- . S . 10S cci E . S S 5- .. ci) S 0 0 100 150 200 250 300 molar volume (cm3/mol) 350 100 150 200 250 300 molar volume (cm3/mol) 350 134 Appendix E. "Environmental stresses and skeletal deformities infishfrom the Willamette River, Oregon ". Reproduced with permission from Environmental Science and Technology 2005, 39( 10), 3495-3506. Copyright 2005 American chemistry Society. 135 '1 &'. "''". 2005, 39. 345m5"3S06 Environmental Stjesses and Skeletal Deformities in Fish from the Willamette River. Oregon eitiI of evidence suggests that parasitic infection, not chemical contaminants, was the primary cause of skeletal deformities observed in River snails. Thus, the' SVillamotte River fish. DANIEl, L. VILLENFUVE.. Intrnductioo MICHAEL Ii. CtJNNtNGfl&M DOUGLAS F .MARKLE, 'the Wihlanit'tte ttae'ruo we.b.': Lit rep. it I iiii' 'itt I American thu 1t14r tlivcrs md e'ejvs's nl.jrc rail It ir square tulle w:iteixliii] ituami ;mtiv',Iijo'r nyu ui ih.,' tJ.S hi llcmwsnurih for tft nuilcs Itnutigh 'fluxed .ugii.,'iiltiir:ul ::iisi urban areas tuItnItill,'I, (i1'.'g..iL'Ctal'19.'sl un5'lrCpohio'anarea,bcIori'oining LAWRENCE Ii Ctt tills, JEFFREY I Jtt FINS,' KARA F. WARNI It, FRED T1LTO1'1, MICIJ.'%t I. L. KENT.! VI ItGI N IA C. tEAT RA F. . 000LALAI SET F1AJINTAN1N, ORAPUIN XRISSANAKRIANGKRAI. thc o:.Ji ulia l0;'s'r tigoiri' I'. 'I'hc\S'IIlainetIe haslitishionni to 10'.. ti 1r1'g.iliians. aid hio'ttltlaiiieno' tatlo'.' Isi neil the ROBERT GROVE, AND mot hiht' pititirtivo' .igrkuttuiat ro'gli.ns In the' PacIfic Ni uhner',t ' ' i is a significant 'tic \Vlttaiuc EUGENE R.JOHNSON,' KIM A , .'tNDERSO2. I )',rin:o'nt ol F :r,:/,:fl.,,/ a.'ui iF'/,r'I( IA' iflus it of Mrr4wOxr. oi:,I I),'pr .0:: :i J I i'iIdI.J. Oovgon 'a.'.' .':.'r..ai 0. or: a 9,' , :a a ingrait'xriidon,nurserr. ttndsp:IwnhuuglutohiLmi lorsalinii, and ne:J'l\'.pi slociLet ottisli xc horn id5'niulirdixo lbs river 3,. tli'i'FC:lili'ilLll Itching is i1lllxr, aid reciiho'uut speLls's are fistird OhiO 'tll)l 'it I tic'. s'ai too thu cturh' 110 is. the .ure'g.':tu 15.1'.. tinciot of En'dromo. mti nec siiptiiio os if sk'tc tat di'forinitk's nh'nt'al ç'u,ih:tv r011 The Willanierte River, one of 14 American Heritape Ricts, fhOiA:S tliioughi the most densely populated and ajri:.ultnrxll' productive reon cf Oropon. Previous hioloqical nloIotorxxJ of the WI axiette River detected ci toted frequencies of sitoletal d omCtios in fish from certain areas of the lower oJIewberg pool INPL rixerwile IRMI 2E-55 and middle oWhoatland Ferr fVFJ, FM 72-74i rover, relative to those in the upper river iCnr.'allis CV1 RM 125-135). The objective of this study was to determine the likely cause of these skeletal cleforntities. In 2092 and 2003. deformity loads in Willaisotte Ro;or fishes were 2-3 times :preater at the NP and WF lo':ati:.ns than at the CV location. There wore some differences in water quality parameters between the NP and CV sites but they did tot readily' explain the difference in deformity loads. Concentrations ofhioavailablu metals were be!:.. ci election limits 10.0-1 ag;' Lf. Concentrations of hioavailahle pot, blorinateci biphenyls )PCBsi and chlorinatnd pesticides worn goiiorzi!k below 0.25 ng/L Concentriotiw of hi':'.i':nilable lsaiv:x:.lic aromatic hydrocarbons were generally less than 5 n;jL. Concentrations of most persistent organic poilataits core below detectron lies its in na r.c: : do tiosue s..amp ls and sediments, arid thcso that 5sero ctiecteci were not significantly different amonrj sites. Bic'.;say cf Willanoette Rwerwater estrants l:r':'.'i':leci no eviclencethzt unidentified compounds or the ortople.n nioture of compounds present in the o'.'trx ould icluc e skeletal cloforniities in cyprinid tisli. Howo':or. eeiaorcarise of a digenean treisatoclew ore directly aw.t toted 'fth a large por:'entage of deformities detected in tcn Vcilltnette P i or fishes,. and similar detoruitties '. re reproduc ccl in I;!'.. 'ratory fathead minnovs exposed to cercaris oyrrri;tx1 from Willantette In tvillaiuieiti' Ricer i'isl's. hii.togical nliillitiring has been vyi,t'xlx' used I.' o'y,ilu;uc .lquatic es:t's''ste'nl health and lli'1iit.i iinpacs 'it anthorupegemiii' aclivilirs 04 has been sil4ge5k'Ll hal skeletal di'tonnhiirs in fish sCulL .15 a usefut hiiiindii',uii' uf Ijiltiti"..........;, and c'iiluiiiiin sk'ls'ltil :l;'firniie',. in iu'i'n0hi' itt has Ito used lit itiotilijif liii' 1 licti populaunos iiIn I 0192-103.1, thstnckhencs' I lske'tt'Lit itoh ritiiii' inntil unit pikemlIin000' iIri'iiri,''iu'ii,io hc,ihthi a0'301't?o,'5l i'iihlc'ctcct from tlii' \cwlii'rg , P1 rcgi:'ul , cx- io'nding horn liver mite htMl 55 to 20.5 Figitri' I, r,nigi'd 1'. i 14'- 13, Nmnth ..ii pike'iniuin in' ske'leiul '''un 24 i 4xt',urttiIv rift. scc're tilso, cli'raied rl5"i(,, in this' ntdctk tVihl,uiiucilr urn ,i.und tiM 12 \Vhi'atlont him'; Figimic ii, Its C',litF,ist, the skch'L,d shcl&rniiit' Cxii's Ut juvenile noruhit'nn plLs'ntiIonn'c' e' .5 'ctd from the' otpper \\:1tlann.lt Ei','c'r tiM 25 r.uy'd loin iii to 51''.', '12-- !Si. Northern piio'mmiiitiiuw ails U' t the timil' sporE's unpacte'it. tiltS species cahlccued Iron liii' Nw'lmc'rg region and associ;mrcd ttibutaries in 4's'. sku'Icitil deli 0 0111'. rats's t'tt'5','ot'd2s''t in tn species 4/J As ii to'1 'Ii', t,i'muo',nit,,ri ig .1 skututal ds'tc'mxocies in tVibt:mwo'LL river hisli supposEd that fish 'ruin Otis' Newherg re5 'mu ;uod ttiiddhe tVihlaiiio'tie l(t'rr hash signiil canthv greater di on oily rates thor Itoh from the upper Vtttajnc'tt Direr. In the nil-tic t i05 Pi'hbtS;ih5 ii tsp the Nes'tbm'rg 'eon of lie' V'iltayi,'ti5' liver as a source ihi'lr king wato:u hit uthan cxpuulsion hieiuhtcuoi'd poibhic cxnc'L'rn rehiic'ib to the' rc'prLs at uti'l mud fish Ii ttp: ,'.;',cicw. IniViuiiCt.i...1110/ safeV.'illcl' ii ,'vs'nlnl:'v'r011mmnme tuir, ' In 1040, liii s"eainphc'. 115% of 11e'OPli' siirvo'vc'd ua.prcoiss'd 'e'xln'unc " c" icons abut the hcvc'l 1)1 tuidld' chui-inicxis in the liver mu.l's'gJu Daily JIino'r2ld lob. 20. tsiolOt, 'ill it research tsas a rcsponse to such CotiCCifls and, especially. scienithc ox i. 'nolan' concerning potential causes, il A with' tarOt 1': ct':muthcih, pho'si rat, and biologurat stressnurs I:,uec Noon fls:,ji'tdLi'd wi Ito ski'ht'tul ilrftrrniths'c In fish. A v,,l'i'l'.' 'b' ctjemiurtilt, miduiding tue'a'w' iiinats, sue'ti as k'ld 'iii ulttulur'itiiis iutiY:'plulcpttluli' pcsliridcs, ao' kaii.no'n Corrxipooiiiauthsorptuiia': 541 .7,7.5yiIfl;5.jr 1511i7:07i In?; o-rnaiL ksoo.and'rso got iats'.t'iii. to induce ne'tlu','ilimusi'uI,' tanitugi' thou can result itt sk'o'tm'ial 0). Chs'nuicals can tulsa cause ski' trial del Uffl0i lid's Curreioi address: US EPA Mid.i:'.muieoit Ecology LaIis.:. Duluth, Mituiesoto. flioparirneni ot Mir.robiolcigv, Otlupanment oil FL u'ries trot Wildlife. drive iuuius'shv tin iui lung mItsuI .11liio'iitilh lit 101555cc wish bane Dopariruent or Envirriioiioonlal tad Malaulir liixiroio4v. lO.lO2iioO4R57O ccc $3725 Publishsd Sri Wsb OOlOtZCOS 2005 cis,,,xe 5,,istv fornatiu 'mu, 'imp nods such as 21t,7.ti-ti'trtuchilornutfisenzo. p.dioxin, p.1 vi'hho,rinuir'd hiplio'nvbo.. toaaphene, and castmiuin bays' b.oe'o'mu reported to sOuse keIntab deiormIl'ks S'OL. 24. NO, 10, 2005 !ENViRQOJMENTALOaENCE & TECHNOLOI3V 3&9 136 N najorthbutary.ntr D Primary study Sites City locations for geographic reference I Dirctlonofflow FIGURE 1. Diagram of rite Willatnette Basis depictinoj the general location anti course cml the Wi)lamemie River and pminuary study locations. thmugh such mechanisms / 0-21). Skeletal rictormitmis have also been linked to water quality problems. iiwltmding ow p1t (25-26) low dissolved oxygen 27-2W. and elevated tentperatures t29-301. Nutntiutsal deficits. panic uiarlv ascor- bic actd amid trvptophan deficiencies, have been linked to skeletal deformitIes in fish (31-32). Inbreeding hasalso hecum shown to cause skeletal deformities, Including scotiosk. lordosis, cut-ved neural spines. timsed vertebrae. and coinpressed vertebrae (33-35). Finally. ttmmmerous infectious biological agents, including viruses, bacteria, and par ashes, have beets reported to cause keietal de[urunitics 36-39). Although the association of fish skeletal deformities with a sslde variety of stressors makes it a useful endpoint (or biological monitoring, the observation ni a high incidence of skeletal deformities, alone, has little diagnostic value. The purpose of this study was to identify or d)agtsoc the causels) of skeletal deforrnitis's associated with 'yVIll;mnie))e Riverfish, with particular ewphasis rn the Newberg region. Skeletal deforniiries in fish collected front the upper. middle, and lower Willanunc River to 2002 and 2003 were cliaracterizedtodetctrnincwhetherreceun prcvalrnccswerestniiimr to thm.se reported previously and to further describe spatIal arid tetnporal patterns. In situ monitoring of river water quality coupled with in situ santpling amid amsatesis of bivavailahle organic cuntantinants mid metal', was used it> compare water quality and potential for direct exposure to knowia chemical contaminants at the Newberg amid Corvallis study sites and determine whether these factors were likely causes of the deformities. Analysis and comparison ol sedlmentsamplesand fish ttsucfrom Newbcrgmwd t.orvaflls was used to evaluate potential iropltic or maternal transfer cii known organic pollutants IPOPsI usa powniirui cause. Bioassay of river water extracts using embryolarval fathead nsinmiows (Pimepitcth'.c prome1al exposed under controlled laboratory conditions was used to evaluate the potential role of ttriknown chemicals or complex chemical mixtures In causing rite skeletal deksrrnities observed iii Willamette River fish. Field-collected fish were exatstincd for parasites, and the association of parasitic infection e.lih skeletal defonnittes was quantified. Finally, cercartae of a trc'tnatode parasite Identified as Apopira/Ius dopt lens) were collected from Wihl.atnrtte River snails (1-iwoin len/a s.rr'#m,s, (lie interttsediaie host for A c/on/cm). and fathead ntinnr,ws 3496 ENV)uONSIENTAL aCIENCE & TECHNOLOGY VOL. 35. NO. 10, 200t v/Crc exposed to the cercarise in the laboratory, together. these components prosided a wcigtmi-of-evidetsce-based, empirical approach to identify the likely cause of skeletal sicfonuu ties in \ViIiucmtettc Riser fishes. fsletliods lJsJ CoIIes'tiu mum stint I )e'kmruiiitv ( liaracle',iirullimn. larval and juvenile tlsh y;rre sampled Irons May to October 2002 and Mayto August 2(511. lists were collected hr beach seine, cast net, and dip net. The three primary sampling areas were Ncwberg pool (NP: JiM ITSSI: lower Willanteitmo, Wheatand Ferry livE; liNt 72 71: middle Wiilrsmeue). andCorvahlis hiM l25-13t3 tipper Mllamnetteu. specittaerms were euthanized with an overdose of MS-222 ltinquel itricairse nwth.mnesulionatei Argent Chemical laboratories, Redmond. WA :11 500 Pl>00 and tixed In 10% bUft5'rt'd fornsaitn Seventeen species were collected, with Puvm'/mocl wi/u.s ois'goncnvls Inoribertu pikensiniouw). Rwimaniaomus baiter/us lredtide shiner. mflo.stonncs nmuermc'/ttnilu.s tlargescale socket), Mi'/rs'/o'i/us eanrinus I lem tithi . ci tad .'tcrom'hetht.c a/muuncetas (chisel- moutlu representing the most commonly collected species total sample sizes 1000) 1/Ui. Spectmcns fixed icir a mInimum of two week>> wereX-raved in a Faxitron MX-2i1 cabinet X-ray machine lining AGFA SiructtirLx li-I t)VET1i Industrial rsdiograph film. IL in was developed ustog a Kodak X-OMAT model MOB developer. 1 5 700 usIa were inspected (or deformities using a 10 lOx ocular over a light table. Presence or absence of 12 different categories of skeletal deformities was scored additional tietailsimi ref/U). and our analyses were based on the number of deformity categories presemit per individual Radiograph>> of deformitY load,i 11 /u pius the number of precatidal deformities. since casmdal deformities mitt' unufuurmis disoribtited ) 10). On the basis of randomree'.'aluatlon o(550 (ISIS, readercrror was not significant Ub. In Situ Waler Quality aim ii Jilt avaIlable ntrmuhmauuts. SEW iJs'seriptiout viii! Saunpin' (o/iertioum, Willmsnst'tte River sampling sites wect' chosen to facilitate tm'estigatlon of sc'asrtnah mind spatial bioavailable contaminant concentrations cit Newberg and Corvallis (Ptgure I). Two sampling stations were located at Newberg, tune itt the south side of the river tiM IT; N 451602' W 12254.OiN) rind (mite a fw miles 137 dtii wrot the nt Is side of the rivet filM 44: N IS 1527 W 122 532iiJ. 1 so liii us wow located SLt (ti rvallis filM i35 Ii N 45 20.15, V 122 .51.00: j2j N .I52..sT. W 1223iL47'i .NLwbun stauuit I tiM IS sees about 25-SO ft froni the soicUise. and 11w l.j-al teaser depth was 27 15. Ii fro' ii die shore, and the irte iwater depth was 20 It. 11th (.orti1Ii statirros were about 15 ft from the share, and tlw I eel water depth Newlaerg statIon 2 IbM 11) was about quantilt: afi n of urganochrloeltics, ongarionll'rogen lristtcidus, :itiI . t'Oitti Iris plt.ltr' ps'stic ties ate' pr 'sided elscs. hit s' till Thu t'lvcrdilc ari'tu;tl Ic hvdros-arho its Pt! I" ontatnimills freeti'ln let'. 'elitnilicle ciIcenuaIcl to - (liii.. 'fill ditt'ctt 'Ii tush qlertuttiari Ii 55,15 pi'rliriiiu'd lilt ii F [PLC: tutu dual di-lecttcar ho lilt:: vs eon' cit Iiride tire'-, hth with I ttaitlliple wat'u'l'rlgtllr. Site lit' recencr' detector had an cxciht:ott w.u'oli'mmgth at 2211 tril cnticsion ss'avelettgihs at till, arid Ito am: il,' lied' ccliv lt:mtI sktcctioim signals tv and 2%) not. I ttts' ti:r'e',' clintpl.utsds Ilnorene, was 7-U ft. Flow near the Cea-s'allt.s stics iii Slav 2tr112 wits -P0Ot) to I 0 000 ui1s andhv late Jute lied deretsdti ft3fs. AL the Newberg cites, flow In S1a 2r't'j2 ss as 17 (Jr.JO itt 'lILt 20ii00 11'!s oral by la:; July i-icasid ii 7111' il/s, lire Coivallisarsta was gencreiiv hllts........12 Itj intl characterized by shallow gravol and srdlntcni riptuip. iti the Ncwherg area, the Willaurutic liver was at tich ii ecp;'r ;0--Or itt, and there were no shallow gravel reds ncr In' s; 'ide sites. The Willarnette River hiss vet-c .roep hairks at ftc cwlrerg area: withIn 5-10 ft of the bank the Liver lt I 5lit ft deep. 'the bottotasof the nyu iii iiiisw' ii IS a urubittati.ii f o:L jial mud. Water satnp1urg was curtdri neil ii oat ho to loS; in and 2003. Thi'u; 2 1-rI satttplittg tV'tltS wre curcrpkhrd per hr di lii' array: the rest were yf,;, r-cte'iI with lit' tluorecetrcc detector. Thu' colunin set) we'. .1 'lieit.'liluctiex Luna Pitt, with '5-tins particle sIze. The itt-il U tnt-ut 5çJ5 run sf111 a constant 11w utile ofti.75 ttttinuitt atil a united gradiens for year, one oacliin app: airnatel:- 5.1:10. 1 ;uiv. and liLly. Sampling evelits were designed to diaracierizr' river conditions during spawning and early developin em. N ulrieu I and waler quality parameters. including dissolved iisvgrn. spvthc conductance, salinity, tots dlssrrl',ed i lids. lctltper:Ltii Lit, J)iI, OPt' foxidatlon reriurti on poten ii al ileptli, a ilunonlum, IlitI its', and turbidity, 0. c-re collected art OIL hourly basis tslth a iSt 13i120 Sonde ifi. i'ellmc Springy. OW Dissolved, btoavailahlc' llrgani ; 1 nfant:ieauts and metals were cotlected 1w dcplovii'tg patsisti si n:liiiig i criers rPM ii and diffusive gradient tltinftliits it)(,t'r in pIJtcL'tivr' irish cages. PSOs COL1MOS d ti neutial lipid Ii -. . tn. dclii enclosed in larilat polymers; :ibing lnvironmuistal Setup lug Techatologlos. SI. I wPii, ML)i I/i live itidisldita] PS1)s and ne n;'r pxsr liese inch Ltd. 11151 ieee mcmii ed ill each cage. Each cage was tisprided with a 'il'.at-;.ii'L-cage-c.ii'lcanchor' arrangatnetit that nsuicd that thy uagew nidsiar at the station and toO ti slav suspen dud tie iLl loin the liverbottomThellvc ISISs isL'LS' IcOn rrrnrpostted tin analysis PSOs and i)G'Fs were kept on mee iii scaled, airtight, atitbet glass cuttlalners dtiring ttattsport to and I torn the field sites. complete I'S!) descriptions have been published 1121 P51)5 weos' gent lv cleaned ci: sediunent or algae abut deployment at the site utilizing .t tub tilled with site water to minimize air exposure. \'i fouling impedance was entphsr ed its eaJunitjtiojis 1 estimated water c'tict'riiratioti, since algae grossih on the deitce-s was till to tnmninial. inifrt:cru it'oo ,t:oe'. PM,ts wci'i' exit acted by hesitito dialyses. inainber glass iUs, Sample vrslorries vote reduced using aTutboviap LV /ZvnIatk Corp tlopietiten. Mi. The samples were then run through gel pemlesition cltr,,rjtlatlgraph IGPC) lenodels SIS pump. 2487 dual ictveiengtli absorbance detection, 71.7 auto-sampler, and inricun rollectorli, Waters. Corp Millard Mi, at id fiactlotiscoir raining ergannchtonintr pesticides. sagan pit osph. ii' pestiekl'-.. 'Snganonitrogenpeoticidru. rt:Iis,atill'Siris v.10 cilk'c-tcd.'lhe Sat. tutu dls4ttvlhrcttzenc GPC calun'inc 5.-ore not eopolvmerparticles. 1 5-urn particle size awl I IiIS_\ Itiresize. The GPC progratn ran with lull;, d ich I 'it iv Italic ai 5.0 1 LI mlimin. Appnopriatelraeu ins were ds'tenttitied hv-ataatralng standards and furtilied ststtspler t. ii. split lItre::; ractions went analyzed using c;i:-dtr:rl-o:ti itl'gLiIrchl.rriite-1), 15(5dual-NIt) ol'ganttnito gull arid organ pltIspliale pesticides) and HPLC- l)At) and Ilnoreseettee flit [sit li: ut.rso\, Agileni Technologies. PalAlto i:5and ibIs' 11110. t"tewlett-Packard. L, I PttlAltrs (Sir. Sanaple maniptilati. tiswei'e ps-rforiited in L'ttltet brown amber or foil-strapped glass c''Itt:Liners to minimize TJV(vis exposure. Detailed analytical tnethods used lot i'. 25 I. c enaplitltdene. anti iitdn it' 11: :',t p rca:. s-tire detected the ttcet.luut rilu'J 'rUt er 'lutin t svslettt tIn: LOtte prognant ran at ii':, aeclunitrile fin 10 tom, was gr:tdu:tllv ramped up to 70% cet'itiitrile fir 15 00 ri. arid tbsetr rattiped tip in 110% tacct'trittitv for if inlet, 'liii' trISIttli sea'- held it ISiS acetotuir: k or miii and then ret::: i'd In lt'iC- atid mimalveed by 1111(5 with diode array detecli in INtl II itd 11 tumescence dets em ion. The approximate reknit 'n 11101's ii ni lii are naplsthiiune, I 6.0: acenaphtbtvk-txe. 16,5: 11w -reLic' 1$ P ithenanhlirene. 18,15: anthracene, 18,6: ihtorattLhene, 19.0: prIetic'. 211.1'): chriene, 2si 4: bs'nzull'd antltr:mcc-rtr'. 22.1; heniotiIitfluoranthene. 24.5. besrzoikilluoriinlhene', 21 8: betshetvoit i 7010 [ISTCIIC 2r I dihuti/oli itletit4sr'5uet5u pert-I tine, 2(1,8: road Indo I 1 .13.t-dipycene 29.2. AflerD(il's wereretniu'ce'datcd in thu-lafrorotomy. thee resin- gel was renies-ed -arid immersed fur 21 h to 1.)) ml. of I NI truce metal guide tritric acid i.E"ir,lter scicittilici. Acetic acid end sodium ,icettuti' went' used itS i}ri'srtpprttng electrolyte, atid the- samples were diluted to all net volltine at 25 tal. sti Lh III-?Itt ciii s-otter, the analysis seas 1st- atiodic stripping cohtarnrtretrv Ast) i'l'raceiletect,Seattle. \VAi.Allgtebwater sautup Ps ice e lii [ci ed rhrrt a 0 15-/ittt nw-tn ht'ane miller prior luinct,ul am uslvscs by .V5'i', \SV wus used to tlrianuft ilte tstetis iprurtel . Iiedttct I rn otet5tltils 5 ';t i' ''cr111 ccl whIt standards each Itfetuil it'sly'd r,:i.'fj:',"t' r 'litIOlt .',i,:. It'Ll, rti, .ini toni-lion blanks lucre its..1111111 etch s.uaillitig cr1-Ill, Field hLlttd PSI)sant pIt's 'vane pencil oral esp setl 11 the stmnisplscru Itiriiig chc'p1-tatett I or ceo v'ry_ I teld blanks were ri ceased atid :inalvzed eactiv as rlCJ'J L.;ed I'S!) samplers. Field ettr,teIt' 'us blanks were ops-rted us lie held and wasltu'd sitttrtlallng th0 piocess of ur't ii''ing thu light se".ilrnet tI or algae on the PilSStl e sampling hcviccs. Satnples eultsitinuilg residues exceedIng ihir' bLinks were coutsideccil pisiuis'e Ion residues, 1ltnsport blank salutes 'sIn' ttiult plied Is the ss'ats'rvuikttnc than' would have be'ui expIsorl to ii' cli with the uther VSI)s, The CorvallIs sue was ds'sigiitsted as a lIvId dupli'arc' s102, Field duplicates repuu'set:ted:':utU if :illsatnplescutllecli:d All l2('samnI'le type's woe inchtideih in '.'.uebt ati,slvtic.th huatcht. 1.ahulurel, nv QC '-..tttil'h's included rr'tgcttt blanks. loctilied saittples. and al's-nature chli]rlieat's. Lacht (II' ti' t's'prcsentesl 5 10% uI il,' intl nuiinbs'r il s.ursiples'ait,ilv.ed iii any given hotels. thc'v were- prepareli ,tmtd ,'utalsved in the' sanie ldshion as the field satmipl's. iii gaul c siatudat I '.i;lteutiSs-rsic-i'. West Chester. tAt crtr'.'v-. 'isle tqliculll\.'oirlposed oh i: 1 standard conconirtit 'to .lttl.l tO i'ltil sttiiiLrrd I_Ole 505,0, Ward 11111. MiSI cutest's ' ', lot tilt auual'.'scs. 581 Al/il,,.'' 'l'lte 11:' irs' and mathematical naodehs requi rush Fir COttilistt is '1 .usalvte seater c'clncentratlons trim the ci tte,'tmirdiioil iii In 1151) lIpid ltase bc'emt described /21. The l,hlowliig 05Jul15 II lr':is tisel.i Lu rilculale the dissolved bIoswatlahlet vttmti'i eousceilttaliOLu. c 5, here c,., Is thu d'c ncenit'ation if ,inal1'e in seater, Is 1/OL. 39, 11/0, 10, tOii,i ENvliRoclMENTALaclEraCs & TECiINCiLOG/. 3497 138 the concentratIon in lipid SP\ll)1 I is he exposure Unit in da;.s Mspei is the 1n.ts A SPNJI) in g, and I ii tiic PSIJ siunpilug rate. ainpliiig rates iI1j fur a large wncs of tIC anti PAll contanhtttjnts [java beeti 1'rci u-h utaNe,hetl, 1he mass ni the rntt,il in tue liCt resin gel 11 was detrtntnt'd mitt the .\.SV qwtntititt n. iii ic nv and niathematical In idik required for Cii ilnatitli Lii jttaI waLer t-uncentrati its from tile cuneentrati n ill the DCT have been previously described i i i. 1 he fr lkwiug equation was used to calculate the labile (bjoasaiiublet water con- N 15 iS .100 \V 22 59 161) - and three samples were' coilected 0 21 .567. \V 127 15111. In 003. Liii ic samples wi-ri- collecteil at Neschet g location N '15 15 57, W 122 5 12 md three samples -tet-re colkicii-'d atCnrvtiilis hicatein N 11 2 0017', tV 122 15-1.12, Sediitteni samples welt cc jt'd ditectta into certiticil I-Client j:trs. transported -ta :c.- the labo: ttsny. and ct-i ed itt - 212 at Coty,tllis Iticotion N 1 I 1 - Until shipped lit-attalesis. m'dluttent santpleswere ixirucied and timiaivi'd Itir 22 i-iilritt:itecl pesiit-iltt-s he ii 511:1 lA milItiA 01i21.i, - ml rEt, igcn.'pltisphtrtms llLstlLuCli-S hr I 7-' NEll, hod 3' 'Itt clttlgetti.-ts by I tittit lit-',-\ ittethod )tutl2 At ii) the ilIEQ l-tiboruti,ra, I'nrtland, tilt, Mills for chlot't- crtttrat.ittn. nated pesticides atid Pitt', u-crc -Ci.33 ,501'Kg wet sat. The Mill, for nitrugen'phcphiiitts pe'stit-ides smas lOsig/Kg wit where C. vet', the eoncentrtU a 1 iuettds in the 1 sl I tNt) eltttionsoiuftoti "'' : was the s [ute nfl 1'-A I added to the resingel, V4 wa the vtlunir- f lisa rsItl gel. itif I sat'. tin elution factor for each mct.il ultecnncentratinnitlte mciii measured by Di t'.i 'hiravailahlc ' watem coliL-entralion) was diaterintited If in tile following equation. sap Skeletal Defortaliko. Illoasotty I. Rli'er Water [xtI'acls. Water samples wear cillect,-0 from tier siudi locations during the summer ti 21012, Sampling sites tnclttded two Newberg locations I NP- N :15 1551; ', W 122 7oil 12 and Al N 15 Ifi 117. 05 122 55,1121, lthtemttitaitl i-tire WI-. N 15 05 117W 123 021>55 arid Comtiis CV N i-i :323387', \V 121 t5,422 I I tneachsamplthgilui-, smnitiiicswil t' collected frotn CV utri tite 1 the oLlser tho-- s.ttttpliug liticatititis At each site, tinee 20-f Dab sampie: were c Ilecied in st,uttiess steel cl,nuantv-rs, Samples sane typE-mills' ci heeled at a depth , Jt\- i)ii) wltcreAçwastitc titti-kitess ofihedillusive gel sf33 tumi Plus the thickness 'tithe filter nietnbrane 10.1; junti, I) was the dlffuslt.itj coefficient liii end in the gel tias depioun cot i. t i ctni I time. and A was lime expisur area i. nIsI.stiI INtl.', In Nitilliet ii Plkcniliin,wt ti iti Il'stte. Notthern piketuitittow I cuw-:lasi 0 -is the sfiecies cli ,sen foraniivsisoftttattriit irtaisier .iPCPs. thee crc Alittttdattt at both stud lc.catiotts, reitmtiveiv easy in c Let, reach moderately large sizes, a tid have large nmunie r Idefurmiltes In the Newberg region lu.l.Adults we: e collected front Ntiwberg iX 15 ILIaC, W la T..i'1:l and (irval1is IN 442820,W 123 1.1.751 iligitre I in -lav-- iiine2iiU2 using a combination it hok and line and ci,.ctr.fisliittg and transported to rite iaboratiry it Ice. \Vi-t'vcigiiis r,tnged froni 375 to 975g. and there was ii 'ignil'ic:ttttdllietulLotn the nican SeCI weight of the fish eoliecteJ from tic siudy sites. Ovajian tIssue and assicialed cvtcs iSIL ieitiiiyed fritin giavid female, using eletin, sail vent-rinsed, dAwi'ti'on ml oteade strean, i IN-c go using a Zvmark 'Itirhi 'yap tI 'the p '.aleoi cotleeutr:ites were tratisferred to autuber glass vials iii P( )Ps. Tv;cn iv- 'iii ci tint i na led peitteides were iluanil I ted bvgaschrumati:igraphv ivitli cC .1 F 'Ii Capture d&iiCtji.tii CICI lt(t)irIc corilitig I Lit tnL-llii ,'icI:l..\ itll)t3) lii stuutiry, Portlatil Liii .1 saentv-eight piivhloiutatcd highenvi iPCBi coutgetlt'rs ore qutittlilied by 1.11 PCI) acciiaiiitg to i.PA method 11(1112 (r)Dt-çt lahorautre, Portlan dt tO .'tdtli lionauly, CrtlicefltrattOt)S 00 seven polechthirinated diLuni.o-p-dioxin IN'[fl)i congenc'rs anti In pulvchlorinated dibeuzofuruns (II P1st uco' guam' hi ht,h tcsuintion (A ("Is Analytical, LirtilsIt Criliinihjti, Canadal, Method detection limits \ifll.s fur cltl.rinafed pectiudes and PC1IS ranged from 2.5 ci 351 /1g. kg wet sat. \ll)Ls fot PCDD and PI73Es ranged itoni ft I tO ii.t 5 rig-Kg iset Vt. PlvcovarLtiv ussue;iiei-te samples Leach Iii ni aseparale fishi were attalc'zetl per stttdv area. hr statistical analysis and pli:ittlng of IIUrcS, o.incentratiouis bet-ac the metharl repi.trting limit IMRIi or d.-iectlun Until were as-, ted to be equal to one-half of the Until. When assuu ipitons of' patansattic statistics stawe met, i-tests were used to test fir differences among sitoly sites. KulinogorovStttlrtiiivs test was used in cases that parametric assttntplions seeje it met. .-tiltilysls t,i PoP, In Sedinient. 1,1 mb samples il surhcial sediment were collected -fr in t-wberg and c5r',aihis sites In 21302, three samples were collected a) Newberg locattn 34 mm DVB-pholiic lijiloseed t't a [tvil-phiillc solid-phase extractioti disk illakct hutid aspeedisk 8072-126, 8)tlit1-05L, I 1, Baker, Phiiltpsbumg, Nil, 1-low titles iscre 15-50 titt,"ittin, following extraction, the disks owe Ailed ttnder cactituhit rtnd stored in airtight o m isittt'rs at 2) 'C met night. To prepare bioassay eunc-ntrtite-s, each disk was cluted three times with 5 ml, of nieth tn' 1. Stethantii elucnts wet,' dttd tsratotv Practice-a- ci-ilified atmletical laisiraloiies hr quanhifk'ttuin of a range I The (10 1 of outer ci illeeted at etti'hi site' trIplicate 20 I. sampiesi wasdisided mItt live 12 l_stths:ttitpit-'slor extraction. Each suiistnmplt seas liltered tinder vocuutrt through a 5-0- i'v passing thrnngh to ci .ltutm ii N-aaSl ). For eltell lte, di ictl nteiltanol elueniswere' pi Ted and t's-aporatd-d -tel :: tnt. ttndt'r until extracted. 1 t etnoved) tindere.ater, in all cast's, ciii m-ciuons were made at least 311 cm I-eS ',' tile curfitce titid at least :io cm above- the sedintent, Stinpis- i-xtractt'tn wits criti:pk-red within 953 Ii of sample couleLtulil. C tools: j.tl aced into certified l-Cliemn jars: LIlA stored at saitiples here 'ILIIIYed to I lb 1. of -A na, and :tnt,tinersieere opened and sealed tall aIr ENJtRONt,IENTALaCIENCE & TEcHNoLOGY . VOL. 35, NO. 10. 2005 and shred it ----------uttmii used Ii bioassay. tithi ugh nut tic tihatishive c tnultuntethiitl procedttres designed fiii lIt,: e'str:tcthaut and analysts of a wide range of Org,hLtui- din iantmnauits in sttri ace u alto- 14'. the extraction poiecdure ilescriicil above was designed to capture it -cnllIcattt cross secttott 0! disotili-ed organic contamtnatits 'Ii g k----t iii the rilige til 1-7p seitht a res-ttlting tnetlumintl CII uli'd-titf ate 'imitable Itt the' in a fathead minnow biotsstn'. I tsittg the extrar-n- ifl in ie'etltlrc described aboi s' i','iIlt ethyl avetat,- is the eluvrit. ,ilss->lvedrcstdues of over 101) currenl- use pesLic ides and tufts tare been recovered and analyzed lie etipildoy 1115 MS Ihsemoko anti Simcittic-lt, pers. intil coin- ntunicatti,ni. In assess extraction i'ft'icience. ri-i-sr water collected at the Ct' star was fortified at hulL'S llg.'l. with chltuullvtitus log K,,, 4,7i, a weii-lonau'n \Viiiantette Ilivet contanunanr, Ethyl accEde extracts Seed' tutiih'ieul by (7,' MS usttag the method oi itsenko and eintonieh, iverage receverv startdartl deviat bitt was UI i .5 it 00 Fathead ninticns's iJ /lalii:t'itl,ci less tlniio2 I hpmmt-hatch i weii'ol:itaiyedlroni Cht-'sapi-.ikc' t:ttilttres lInes. VAI. 1_at's-al fatltead mintoows F11s1: 24 lOt it past-li:-tteit , seem e rtndriittiv assign-ti ti lt){l-mt hemikets contaiitittg 111,1 uttt of uk-chlorittated tap water I this i. Each bcsiki'r seas slicked witlt it :10 l.treal II tM Eeakt-t se-crc then randuwir assigned let tile of eight I reahmhhi:nl groups I reaitnent gt outs for the stttd V er enijtj'ul till", _llt'l ml ii dto ) soli Lilt Lottirol 150 0 (1 ,°, 139 Me011 In dlwt; fiX-, IX-, and 1X-Corvahis: OX-. IX-, and 1X- Irish were delis em i'd iii. 3-7 day old and were InItially fed NP AL cit WI. :X. IX. and IX paraitt'dmmmtt ettmlmtimes umitil ab itt 1)1-14 days old, thenwere switched curt mitixcure ct brine sltrimttp Illltllrthlti 1CM. i.lrine pescirt the vciumL: i the apprpriatc:slruct thcsolved in 200 ml. dlii Li. provIde a conceotralioct erlun aleni tollS I. Ii JO. dlii! I lf,. respeci v 01 river waler cincin traiton Lit the curs et s ci ntit uctt is, assuming 1006 reel series. Mitliatti 1 tess ddcd ti. eadi 1 the 4X and ix lrcrittnelii uclt that the total Me..)!! idirititp rind In cwe-drr',l .spiruliitrtmlgrtt iAIgit' 13'asti__kfler rrtu1 )-i fish ',-cue titc'ti ted TclirsSlttt flake hod urcirri Sales, Blaekslsmtrg. \\i. Liii, 0 05! tell: sort us lCerc ci 'Ilected Irrim the \ewlterg concentralion was equierilet ii (i chat 01 the L&' reutlItdHIC and Sc .0dFt1IV CidCl a k { liii test si.luti II was rriiereed dailyby drawing the iriltulit d en to Iii ri. rind addiiitr 100 ml. of fresh tet solittin cuntatniog ni.ininiil coilce,ltru- rtrt',t ligiare Ii Irons iutie to .kicgust 21.ttiR. Cc'rcrtrlae consistent wIth those dcscmlbed he Niettil acts! Macs ItO, ipigure 2a1 tlonsofcxtracr, iii I:enr or both:! ho locauin .1 oalt heLr on the exposure hncit was assigned rruidinlv titI ill beakers were aeratci thtrtnghout the 0-epositre iluittl ir. tnrmmmlnmssein studies, larval frmthertri ni intl uws After 5 days of exposuic. survivingtish were entitled itid transferred 10 I -L plastic containers [or gru-ilt i -d tb 25-30 post-hatch. During grow-out. itsit seer iiritttainied In dtw supplied from a flow-through system. T0r.iiigh err both exposure and grow-nut writet lent per,ttui err maintained at 21-26 C phulipetlel was Is Ii li)hi. core Itars'ested fiotn inrliviulurtl tiall be holding snails itt tsiluIi eiitm 2.1-well tissue cttit tire pit 's Iti 2 inc_ui water, For 1 varvirtg age 15110 ii were exptmed Ii kiti'';n 'liii entice rhts ofcercarlae ci nil iSs ate! Ijtllt,tI I nials with s'erv ytttmttg fish testtlisd in ItigIt tiiitrtilmly In e'xllisstd [kit able Ic lit-i deoce itt tnlcctton, t'crtehtal defurmittt ms and associatits of wortats with dirtorim I irrnr seas determined by exritiiiltrtt ho ofwlaolc', pteserved isil that score cleared with ii'psin rind stained with with allan joe and olierrmn-red S ;iii. Fish were collecrett rut ehtimci (I Ii daik, and FilM were fed S/nrarnr.l )Algrtc Fern I.:trthrirre. Petaitima, c/si cult-c daily and brirte shriuip n:ritpltl t SI urine Shrimp. Ogden UT rice drill., Ltisseiived oxygen plc ainnionia, turl ttiuritr' were rraoniLured daily. Atthe end ot Iltegrw-iulperiud, fishitona each cuntaitter were te-ansferrird 1 a - cm-diameter plastic tube with [toe mesh at one end PVi Insert), The etictre hatch if live fisit was immersed in a (1.2... calcein Siptita C-iii17: St. Louts, Wit sitctin pl 7)1, stdtLtcd lit to mitt . trmiits[etrd to clean teater liii iii olin I. distrilmi, sit! mIro euthanjzod hr irtqui1, AigenI. ltetli.nrcid V.\ I ro h:tnizeil specinhsits were inmrnedialely exaciticted hs lire i issetiri tlttcrr seupy usmg a Leica MZFI.l Ii intmurrsiun in ri 20)1 nuil Lsohmtii.n ut M's-222 dissecurri ntierilsdi..pe ithiutit's rtitd Stout. liellevue, WA equipp'rl with rt it ercitrv snip and fluot escein.igrecn fluorescence l.rnletn ititot. Cakcin staining allows fur dtrect vlsutticzatiort of calcified skeleiai slrimctitres Ii). Fach specimen was eLmitned fur skeietril del entities, inriuiiicig smut- sls lordosk, liwed vcrtchrae, compressed centia, cr1 raur missing sinuos, etc. Seri'enitmg ofst'v'r il hundred fish is part of assay ds'vJopiitetir rintirnied that all these types of deforntilies ...crc detectable he this method, Vertebral developitietilsrasrtls:swreduo rmscaheuft-5usingaeriteria duilnid li this tuth'. Digital imagEs of each fish and closeups ol delurmiltes. if detected, were captured rtnd archived usinglmagerrut puts t 3.i i.5redlan tnrnoiicc, silversprings, MD). in scene cases. exiimttictaiiins were spread irer 2-3 d. ItepIiCttes exsunincii erich day were selected rttwlunily. Survival In cite nj of crposurc d pusc-hrttcit rtrvivrtl toexaminatlon 21:- nd pust-ltrttc)r,attil pr-rcurtinf situ tug fish whit a keietrtl (letoemile were del :1! iii I jr each replicate. Deveinpinetital SI mc disleibuti irs ci e determined fir each trcetmnwtil. (tue-way analysts .1 vtriattL.....is used. to test kic differences in survival r iticietetiec of deformities alitotig Ireattrii'tsts. 'i i enpriritmel nc Km itskaiiWallis Lest....ii rrink: was iit-i1 i. test U e,rlcnemil-tclrmtcrt dilitirences in c!es'eh pnmecictl s Iv disltibutione. Skelcial Iie1i.i'iiilths ItLias.sav Il_I xposuretnt;iopltsiffos ddith'tcc Cerearlac. rhrmrrcicrizrtti mt of prtrasite association with vertebral drtoututies iii Wlilamette Ill--er 1ihes was based on exrtlttittati,mm of iii stutogicrti seclions of fotmalittpreserved fudi ,is well -is whole mlitictts if Irvlisin-ckai ed ii Ti) driv', ptmstex- Psili' ttli'riitillslt were placid icc rt Pert dish, coveted ssitic '-'ccci tt roil ex:imuincd at 25 sr Ii l;tshi were also ev..ilurmte'd hg radirprrphv, as desctibeml liv %Iartl....I al, 1)4). ReuIts and Discussion 1I'formlly inrids lit VflIriiaie1te RIt'el' libili. One difficulty assiciated whir the use of [isis skoltal dcfortartttes as a bimntutnituring toil is cite ittek-. of .htitiurinrttlomi on normai Isaekgrt'und deformity rates One stttetv In salcnotalds suggester! thaI 2-- 5r' ion' he a n'iiitrtl limickgr'imndrate In wild p ptiirtti mis s ticwer Cr, it is tmiiclr'rir tolltecher deformity rates differ rttniing species r.riii' ii!. hiolttlrtthOtts ssiihiti ratrs are itsually hiiottinit.ttiisg rtpptiirrehes b,tscd oii skeletal species. l!et'rtusc backgrur mitci.l dcli itO utikmin'e'rt. dm'iirntill..c relvi ii Letitllirrth crsprttirml cotliltanisotts, ouch as s-ear ms vent cltniti,mies, ir ci etlpntns-.otms hst'sce'ti locations with slitmilar habitat, ehitttiIc, etc. itt t\itluti'ie'ltc illvcr bisla, the 11 jrteked go 'graphic disipari i- its lreqttencs if skeletal delorinntics suggested oOzed ' 'bLots iii Neiehcrg aitd Wheatl,ttirl i-u I S)ttt 2lti)2--2uii.i_t :siiit', is'ue citliseSilt wsih pres-liuits sntiltes iItii rep rt'il t nrc Itor lctciilcncr cf sk'k'tai cti'furittities In IVi lI,tnieite' Ills-er fish front Nc's-berg atid \Vhemttlattd Few rektmive ci U.lrv,uliis )J/ !l.tttsctg the live species rn si ettimi liii 'tmiv satutp led. pt cent ireqtmettcv if dcl ,rcnt lies torts grttei all" 2-- tunics greater it Newberg rind \Vhcalland iii rr unit rtt t it ratlts l'rmhk- Ii, liii' univ exception torts the l:im ge-scat' stt.'k,'m if rtrluJ.'Imo tiSl,''i'uule'f,5r1.i , s-;lttelt torn mitt, only c'rit snytnid of ihe fne twist cirmimnirtiv collected kIte', \mti.ttg lime cvpnctids. iuentn dcliitiititu' lurstis were usiiall sigimitteantiv lowet for lTurrnlhns fish rho fr fish frirtit \VIre,ucland Ferryir Nes'berg lJtrble I Thr'-m were tilt obvious geographIc or lutf'4irit iille-renee-s tim at explained the differ- etices observed i Irti. merrill, bliutttniti.'ritsg il skeletal deformities in fish collected at mhfferent I icatii.urns along the Willntmiiette lth'cr itt 2002-- 2ei;.t sttpported the clrrclrmsmort that fish ppulati its ttertr Xs'scberg and Wheatland Fctmv were note likely or hrilyc skeletal defom httes than fish irons Cumvaihis. Spit Itmsl ricrtl snmitipht's front inttsetmm collections cimtcuirtttlugit pr crnidrml tloirniitv loath ttt,33- 1 4 perfish) Itt liii) mt rscv:li.-tg amol WIce,itlattd Fcrr-, a Iossr load itt alcian blue cold aiiaarirtrrJS-viained Ilsit i IT. The tnthosls 1. I/i rut \simeritlrictd 1-err., utnd mi lower lied upstt'eant ol Cors'rulhis ict 15)7 Ii 12 .1 )1 These ltist'rlcctl data do trot andstatisticmti anribsis used for theparastle chtarrtctcntzannmt 11111) were reported elsetisere 47). 1or iabormttorr tr'jlsrrrlssittti stmtdtes, laltatrs-reared [atIic-td ntinnnws va'rcIir,.iitied lenin Cheastpealrc i :ultttrcs [-layes, \3I. Ulsit were lielii itt dt'chui'rimiated tap wrtter i23-21 C) to ensure ttnoxpised fish did not become hrth'cted I-wit were maintaittedin stril re is rime nquarlawlih htolt.iglcal illtcrs 11152 In dlstIttgumsit slung three possibilitIes: lii site ,.-rtnirition. and the range ot sciriation we delict Is normal; 12) talc's at 0t i-allis represetti time hackgrtamil. utnd Newb.erg rates rice eli'vtic-d: rtt 0 rtmc'srit Net' htirg represent the backgtuund, rititi t.;itvrillis liii ale' depressed. 1 Wmiter Qtimilli v (.Iirirrii'ieri/,ithsn. \cutmmiottiil. nitrmtie., piJ tetstpe'rature dissi lye! ox'gets, oxldrtttve ceducrtsutspoterttlal., VOL. 35, tiC, tO. lOtS) ENVtfiOtIMEntTAL 3c,iESCE & TECJINULOCY. a&aa 140 L.. FIGURE 2. A. donicz,s infections in laboratoty-reared fathead miiinows A. Pa,aplophocioco:is cercaria unto F. neos used in exposure trials, Bar . 50 pot. 8f. Metacercariae lairowsI associated with skeletal delorinities in cleated fish. Bar 200 pin. B. Metacercariae iii surrounded by liony prolilration at base of a vetlehra C. Metacetcatia in reyion of severe lorilosis. 0. Dysplastic sertebtae and lordosis a; site of infection. E. Meracercariae associated with lesions: 1 and 3, no sigirificant changes; 2, dyspiastic and broken spines: 4, severe lordosis; 5, metacercariae in base of anal fin with is changes. TABLE I. Frequency of Occintetice of Piecaudal Deformities and Average Delonirity oaitv mi the Five Species fiosi Coiurnoiily Collected horn Three Willaritetre Rivet Study Areas in 20022003 Ptydhedosoro.qorrenais, northern pikriminnow P I m/'s'rao Laura rtus redskIn shiner catosromous rnacrocheiis,,a. iarçiescale sucker 23.6 0.37 14 2 024 <-0029 8 14.8 0.21 . 0016, A 1394 Mylocheilua c.aurinuo, pea inmith Acrc ra lj 'i(uraceue chistslmoutir 14.1 0.20 . 0.032, AS 305 24.2 0.34 a. 0.033, B 30 6 070 2h- r1 0 01 Off' Nt 6.9 0.09 ± 0.01, B 928 C ij 33 938 B 4 17.3 0.22 0080, A 127 Off DL° 0.016, A Corvallis Whearland Ferry Newtrery pool species 2314 c 22.8 0,35 1 U 21.3 0.34 20 0 O' 0.021, A 1205 001 os 1001 0.1 11, A 47 442 9,4 0.10 a 0.038. A 96 14 0 00 C 251 " U 10 B lOo 1 U1 Fiequsncs' :4 'wcurrerics at precauclal ciefc'rinitiss. Deloriiiity load.: ritiniher of detornii cnte9orues present per individijal; newt ± SE (SE is incii'oiclual ratherthan poc'lsct(. A.B.0 indicate siyniticant difference :etn'sYa rite. p .c 5,95/ brired rn Bonfrroni multiple ranys toot. 0 Carrtplt sire. and specific conductance data were collected hourly during all 21-d sampling events. At each station, some parameters showed strong temporal and spatial variation, while othei's did not. Diurnal temperature variation was greater at the Corvallis sites ii 2 'Ci, than at Newboog CI C or level. Temperature patterns were the sonic for both wars and worn' generally 12 ± 1 C In May and rose to ".22C :b 2 'C by the have bec'n linked to skeletal defomuitlt'o in stickers 125-261. bitt we are not aware of ank' reports that link p11 7,2-8.11 cuiiditions to skeletal det'orntities in fish, 'the oxidation reduction poieutiai cOBl'i probe tended to tirill tiler abotti 7 tO days of depk'ivntirut. Sitiec till deployI iir'ii lower e 21 days. data alter 7 c1ar went' exclu tIed from Ihe analysts. OiiPwas consLsteittivlowerat the ewbergsItes. end of the sampling period. Maimum Pt! range doting 21 It, usually tn late May to early June was L2 to 711 at the as cotnparn'd to the Corvallis sites, The difference ranged rum ' i0'y.toa factnrof2.Thure was no apparent rteasonalitv Newberg sites rind 7.2 to liii at the Corvallissttev:t'hn'sefarge diurnal p1-I changes were seen in both years. At all sites, ii the Corvallis sires and liii It' di ffert'iic e between 211)2 arid 21303.The[e wassonte evIdence that the OflPlncreasedduring therewere sinai! decreases in pit from 21)02 to 2003. Nlglittinre tits' season and differed among tears at the Newberg sites. pus were similar, but the daytime pit showed a strong Tljr' (188 value is a direct reading of the activIty of oxidizing geographic diffcrence, sdth the Corvallis sites often l and reducing agents in the water, as they correspond to pu wilts higher than the Newbergsltc's. Low pH conditions l 3500 EN VIRONMENT.AL OCIENCE & TECHNOLOGY/VOL 30, NO. tO, 2506 oxi dat ion redsrrtiun reactions 1521. In general, the Corvallis 141 TABLE 2. Meai Coinennations n)I) ol Cloatailable Orgaiucs Estimated Imoin Coiiceutmaiioiis Accuiiiiilated in Passive Sampling Devices Exposeil lot 21 d at Two Sites (2 locations pet site) along time Willainene Iliver' Cotvlrn Nowberq pool 1PAH" 3.17 2 147 nrithracenu" 1.32 0 0.92 plinnthrnu 1.22 1. ('.722 i ('.6(0 '.hlrpyritrs 0.94 ,, 0 1 97 :641:1)2 0.61:00.07 9(1 1 (.923 ('IS. 0 .0 13 .. 0. (19. ((:74 ii5 ".6' 6' :1.049 roil 0.01: ..o 0.6)2 0 .323 61,900 0.023 0.007 1 3:19 (ll,,l.:Irinn . ' (1 0.43 .t 0.12 .46' OPT p,'." DOT' 1.72 ((33 ±0.27 0.9') 10 2003 0.34 2.01 :,"3 0.53 flit ornFnh1uII)-" 1PCEI1' 2007 2903 2002 0.609 NA ('.74 or, 61.011 3. 6.202 ariA (1.021 00 (.33 NA '7.01 ('.606 0.) 1017.764 ':625 0.769 ,'iIli ':":'ii':'ritIiti'..Io. -' dnluLicin Iin'iit not nh...'.',ii. C",., SuI:.I'....l'tjnu Irifol' 101011 lou 'ornpk'L. Ii'l .1 nrioIti.'r. Ojnificnnt diffornnct L),''nuwl silos :5111 sooir. Oiauiificnjil clIffururlcl' lOIS, .011 20:3 :1113, "tql'li{lcOl'lt ffnrsncu l:01'.,l".sfl nitu's. 7(0311(3. Tnrtj-ot rtnnl'11.',.t. TABLE 3. Concentratloims of Cldonnated Pesticides, Polychl ominated Bijdi en yl s, P olychlominaleil Dihenzo-p -diositi s and Polychiorinateil Di hetioImnans Dete cle ml in Oocyt Ovamy Tissue hum Nomthietmi P ikenmmummow )Ptychochemlus orgeotiensmsl Collected from rJewhiemg Pool (NP) and Corvallis (CV) Study Sites SE (tig(" ' cornpolntd mediiut 015/50' NP atidrin 4,4L000 4,4'DDE P50612,43 3234 i' 1.62 3.1:3 ± 1.34 4(00 . 3.2') 12.4 1.74 NP CV P '1.60 1.62 (1.191 1.69 35.0 1.60 1.62 3,171 31.0 7.70 0.942 0.144 1.69 1,65 1.62 1.62 02399 P5018 12.2',91 P00191 12,2'.4,O,E.i (1,7 P0611') 123 3'4''3l ('.49; 1.65 ('.96 1.65 1.65 4.20 1.65 4.16 PC:E.111:l2.3'.4 401 P5.0130122 344 0'] 2.43' 2.3:1 F...E1 l,l2244 r 4 TE0ipanp,sI 0.04 i 1.27 A (01:74 0,671 3974 0.163 0.999 1232 1.05 0.67 0.99 (1.213 0 .46' 3 i3J'(t soit '1.1 Fur Ifs purposes of cnlcitlstin,j rt'twns, nwrhnria. and statiotics, ON I blot '.. '.vors as'.uniud to, On 51111)4 Ii) '2 tli ninililod d10nctiur'i loOt. All C nawttrati 15 rn(YI1l Ill 13 01 ' Irillate sa 1155 1 "a ," I TOO 'bit 9 ,ii I ,. rtd a t in Ii hl us 5511 'Ir mn lj,tn I than the MDL. I TABLE 4. kmcideiice ot Veitehi oh C eloitmimiles and Wetac.eicam iae in Fathe ad Minnows (Pimephiales piomoelas) Eapused to Cetcamia e ol .4pophallus doniczis eortcti of ettpouure triI oc, ae ol ettponwe ce,-eeiia&iith yn( tlaa titiwittitien pleoposie mititlisi when exemitteil exataeti 09 90 11 1 30 8 10 0 )4 0 19 d Th7 14 20 0 8 76' 18 2 30 30 0 4 S 31) 4090 0 5 7(1 17 24 70 "Ii 14 1124 71) a:oo,l total t.:.t.tl c.c.nti'l' 21 19 70 99 0 12 4 1: 1.196' I 1 pannaitea. N 1.0 9:10(99' 911 /82) I o 14 3, 0 0 1'S')) 1 1 1 i'23'.'l.' 'awLs osic lItre cixidizing, orh':,s iuduottg. than Noa'lcrg very (1 uil.i: .1 tb1 & 3.t'v;iLl Is I 1 0 04 91 4 0 auseciated with 0 1 I dc, NA NA 0 2027 NA 171' ion' 0 12 0 (1 (1 131403' 14:14 /100) ns'aLtirs. ('iFOP I'. known LI litllu'ti'' ttbl'ui,I.lbaI gruvo'Jt. bitt it 'is not clear wh1h er lie dill ci cot.' (It 03)111 4'survcd sci old make' Nunclit'rgfish rn ru vulni'j ,ibk' 10 II1l'L'tl' In liv miii ulii,il agents ot' mcnre stuiuptilile I''. ilcl'o'tntiti''. Specific condue't'isiiv (SI : IIIIIIIIthIItCC 911 232) 0 iitfesied % 911 (72) 7 10 12 1 '. worms anoeiaIed with dafermitiea. N delsottetl 4.5 40 0 2.9 '3 314 NA 31.33231' 4' '0 12(971 NA 3064 iE'31 15 08 11:5 4 6' °: 17 NA 60) 9 int'i'easi' in :ul'Itnl''nibt l'nitt 0.113 10 i.29 mg/i. was measured at III., lIt siLC'n. 'aitil t3i'vaIIIs g'iwt'ltlIv havinglaiglier ammonia than \1't'bei'g. '1111' niU'ati" glob' wan ott' robust. In 201)2, dciii gc'Itr'rallv ,ii'rttrted within 2.1 Ii of dc'pli.'vtnrn&. In 2003, Ihepruho l'ai]cdwllhlna fu;'h"strsal' di'plusntuttt.i'helimited data itIdlilttcd that nitrate Increased frito ,luv to Jttiv. and andNt'v.'hrrgnitts. with hrltllsltl01119 .1 sJilfhI iue'i''aae it'' in inlay tci July. The SI' pIIl'r"rn win sitn,l.iu (si built 'i'ItS. 'Fit" dissolved oxygen II it'll 11:111cm 'so,. 1itiiiliit 111 1,11th itt'5. A diurnal pattern wts'tpp:Irunt, u 1 'a sliglI I 1ls't'rl'On' in DO concetitrath Its wore ltbphi'r at 11w 7'wl.t'i'gn1 15,115 c'iiugarcd was seen frcim Moot,, lull' In bath 2002 and 2(111.1, A seasonal cmadtwu'ity slurs hikclv causes 0 the diflbre'itt dcli nitty to thu Curvallis silcs...11 WillIe. O'ateI' tiiililv Ili1)nil'oring guI 'sid1'l ti. ci inpelhi tip i'vldt'nco to Sllggl'nt that dilk't't'twt's It't. ttl.ttt'tCtIt CIlLic'itt 101bl''Its, 111 I. te'mprruiurc'. I >0. or Spc'LIlie VOt. 39, NO. 19. 2905 EN"IHONtlENTAL SiltricE TECHNl3L0G'i' 3501 142 loads observed at 'wberg vet sos Corvallis, hut futiher itivestigatlonofa p itrIellnkbctv,ertn lltPdifientrnccsand susccptllilin to ii if ret i,-ii clay be rrantrd. ltlivalltdtlt' ull tiltiants. J5?.uiCt51i'. tvfl2ttti lit/rn- ciii?ti.4!Ls. ri at bit zn ailable Pt tic v. re it sc at alt locations and for all sampling events igene utile I iigl .1. the 10 PAils uwistired, 13 PAItttccere below ileteetlin limits p ., 151)15; 2003 p .. 0110.t. Itowevee, the' hi tttvat able ctuncentt etuons tuf diel.drin were very tow, It 21 tg.'L, and Iltete was urn significant difference I.tetwct'n vials iNeteberg ii') t; Cuevai)i- p 011113, 'table 21. 'I'he diekhaia conC'entrat)oiis iinwct ed wire anoLuad 2, tilt-fold Lx'low time ireshscaierCi.t',ui ii.utS6og,i. ).Sii and tire well hr'lowthotse. SO. \Vc' are nut generally thoiigtst ta be Susie to tts,h N 0.1 ng/Lt in J' '112, and 11) PA! Is nere belt sc iii 20.2.. I'hrce aware cf any repair lhitkirtg below nato grani-per-Iiter PAIlsphenaatltrciw. fluorantlien i'. and iii hi racttewCi e detected at sites ut 2002 atid 1' At ii .rvallis sttet concentrations a! dirtdutn to ck'hc'ttil Jet urutuities, Chlurpvrhflts seas the utnit sgtitplt sphaie pesticide detected, Dini nthoate, diazinoti , anti aitutphttts-ntethyl were' below dementiout limits ii all Silt's itur till sautpling events idetci'ti itt bum lie were cci! twa nil at 2. 2. and 2 nghl., respt....livelyt. lii 1111.1:',, Ihi' t'ctintzited, biivav;thl,ihlc. water ctncc' ntrauitn for rhl'irpvrifns a'.'ergnd 0.71 (I St lit Corvallis ,itd 1:21 135 up;!. it t",''lterg If attic 2', 't'hszte was little dmlfeeenre between the -a cs ill I Li iii' ,titd Jtth', but there was .0 cue dhifcinit cc hctwr tit the' Sites lit ttty, with 1 lahensiithrcns' utd atuhraee it 1 tnt' equath abundant In 20112, whereas tittoranflieni' was cunewhat Iss abundant (Table 21. 0! the three CU is tkiecic'd at the Newbtrg cite in 2002. phenantlsteni' was thme itmt,.t abundant çl'ahle 2'. In 2003, anthract'ne see'. oust alattudant at Curvaflic, and phenauzdu me was in '.1 ,ihitttdaui at Newlstzr/r 1 the I' ltidi1dttal l)[O.tV1LilthIt' U Is, Jfl 21:02, the Corral Ii', cites cnnsittt'ntiy hd higher total Psi I 21'Alll cttnceitlralions. as cttmparsd la the 'lt''t'tt''i p siim- 'fabh' 21 lIt' and ipt PAl I .:.t.2 ugh. ti Liyallts, whir ta' the Nt'sebetg siteshad 2PM I ciiticentraiions in 2002 of tb ti! 2 ng L In isa 2002 v..is 2003, this ptnit'rit was again upheld iTahic i'. The Corvallis stations 11 raid 21 had tAt Is of:) aid 22 ugh. tecpcUvei the Newberg statints it and 2 had l'A1ts of 2 OgiI_ Phenanthrsttss produces spi it. 1 cur-el Ott' in erbiafish cmboos at contcuir,tuistns near smiler calurtuirn 1.25 alp..'1..1 (53. With low cotts-enlr,utions detected ii ho iii ci its. I 'At Is werenntconsidc'rc iii lie likely contribut i$ the ihitieri'ttce in deformity Irads assuriateci wtih the Iwtl areas. Po!ve!ih.,iniitet! l.ztt):a't:'/.s Pl.M.s: PCB analysts lot this etudvwas based on a sritgi'iini-speclllc approach; however, concenteations of all Cut igennus set-tm quite litw, so interprelation of results 1/ ti.ised in t ltd Ii 1! c iicentratlons. The total hioavallable 'Ct', iritcent t:ol ns were getintmtily vecylow, <003 ngL. at all shies, and nianv sites mute below detection limits 101101 tiguLt 1.'ihbc.ii. l)uring2't.12 ,ind 2t.ui toralbicaavallablei'Cilswvrc gencrahl'cgietsit'r at the \rwht'rg sites h'rable 2), f1ows''r, since tnt st a! the data 'mere below or near detection limits. ii teas hiflictil 1 5 ii raw ani fIrm concttisianis, POt roncentratirns were a [proximately 500fold lower than the clii roe anihsieni scaler qtiztlitr ci iterla, CCC) of{i.ttI4pg/L .5/1 and liter lid rut re.idll explain the difference in cieftinnily loads tin op sites. Pect'ridt's The bictavatiahie 21 ill stint oft' p-DDT,p,1CODD, and i'.j)'-DDEj COnir'lllr,tUuils welt' how igeneraily <0.07 np'!.) at all sites in 2' '12 and Air's . [able 2/. 2Ltllt coflcefltratii:ittsws're'-.- 10-flit I-mS te 11w..u'shwatet CCC. (LOU! sg/l. i.S.O. The dilts'rc'ni-i's ci 31 t)f between estterg nd Corvallis slit's 201,2 to ... lit e iii it dOt; 0.005) weee mutt sigittticztnt lIable 2. tntett,,,i,ti,tt ditimi cures in . 'rIO Nev;hvrg; ltD I' sans rtgstalinttc were itt-I sigiuitictitl it, fir... 0,2-I Cuirvallid. In 21.012, tilt [lIlt' pnufi2: toisdiiiiititd b tl,1.t-t000, (ollowel liv lit itt raid l'it)T lahhe S ,m,t;'iithsl be expected from alder lu:ptisits. I iu,stdvrr, ii, S''2, 0017 domino Lcd, folluteed liv iii 13) at) 1 'III table 21, DOt.) and D011 cr,Ltcs'ttiraiiutis d'iei'ieil tit \m-,'herii SI icc in 2112 and 200.3 were not sigiiitit'aritiv tihlereni p kilo DDE -- concenirationshutii':il 101 a rtci'ntrttiailsat i .,',-.tlll'ssltes Increased slgnili,tattllv hetweeit dtt.i2 ttl hull p (hilt! DDE,; t .1 sn.oi . \iets tiler cu tscenitati' as at 2ewhwg - did not hticicase cigniticatiuly beimu'enii 21,12 raid 201%) 1) I; Table di, but iniriti biozivallable DOT dtncted at Corvallis increased .4-fold up (,0q4; Table 2i, The cause of the Increased pp-DO...concentration was nut clear, but the results suggest possible new iiiptits or sedintent disruption remobilizing DOT, Eteerall, the crrncentration dibfc'rences observed were vet" sttsat t '-'51.00(3 ng/Li, so cuitrlurlons should be drawn prudent lv, Dieldrin cotwetttratiu,t is ccii' slightly higher at the Newberg SItes, as compared to the Conallis sites 3 able 2; 2002 3502 EttVtROt'ISIttITAL CCiENCE &TECHNOLcGY 'CDL. 15, Nc. tO. 2005 h 1 -hih.rjuvrlfus concenlittuu,utus at time' Nu'tein'rg slit's bight....than ut hut' i :nurvziihs sites, however, time cli fti'ne'tict' meOs 1101 significant p - it SIt, 'lii' luioavaiiablc' c'hlurpca'ifius t'inceuiti'atiuns tabsciveil in this studs- were below 11w' chronIc aquatic life cetterla ciflltlluplaisct,..elllnl,wclzimcLi1kziuhotlS reported to be R.celc to lisli 2. listVt'i1 ehhrurpmi il-as concentrations tire also welt below the OS'S, n.uufi'Jmncc' lower limit ofthc' henchunuai'k ct.tnceouluaticlum cdliii ale' iii 'nut 5tg/[. resulting 111-2,5% bramnacetvlcholtnesterase rAChI) Inhibition itisteellucadtrotit L2ti IiiainlUaEusconsidered astiscissiiis'e indicat,ur of sublethal c'fiect AdditIonally, we tire lii1tt5't1c' of an': oi'ei -reviewed reports linking lesser conrenloali inS it chiiirpvtilos to skeletal deforitiltiec in fish, .lE'utds tiioavaulahle heave insital satupleut were collected using diffusive gel thin fIlms )D(Sl'j, iund samples were depltuvcui lii lit-' sztiuie' i,ttsnneras ii coiher PSU5, iiloavailtuhtle zinc, eadittri,m, lead, anti ripper were sletc'rmi-nedfor all six s,iiflp ling events, liar. c,utliuufutn, le,wi , copper, aiud ztrsetlic 111 Otis' de'ic'rtniiurd iii littet cii gt-,uh Stahl samples pulled duiltig tIn' 21111', stniplatg dc'phiuvin i'tmls. 'fhc' iuiu ',t'u',iilthic metal u: ncc-utt,ttiuuit. 'acre - ti tht- mist pert, hs'heos'dctectton timus 'flue univ exceptions tieR' delectiuc tI, lliuutg.-'l. i Zn in a sIiupin suutiltle traitt Ni'wbt'rg and 7 0("l. ii lIt lit a siiglt'"aitiplt'tl,ittt I ii''.ilhiS. Nt-itlut'riltIiL'st-dc-it'i'tlotusmst'ue rcpli'tirtl. zini till u'ilucrsaisihiht's Ii an I' iti stit' wire li'lrutc detection hitnitc.suu built delicti 'ncwt're ronsirtc'ied ;urufiacls, The i esults prts'idesh no evitleiucc that hioavuul able heave musetals 5'S'te'',i likely e,imts;' Itt the difference' tim rlelitrtttitv ioztds at the I sea sizes %I,itcrnal Traiusfs'i' sif POPS. (:rmcenlrat-tono of POPs detected in Wi 111111 mcI Ic iluvet northern itlkentitatmow ovary! oocete tissue wi-re rm'lamtvely loss', ('huitritialed peslicitle concentraul sits 'Crept' tii'rahlv '-3,3 rtgg wet wt, amtml ml" It of the 21 dtlfc'rtmmi eliltrittal,'mt pesticide residues aittmlvi.t'd vine detrcicd )'i1mble St. I if lie three, 1, 1'-! 1110, was uht'iuzctert with [lii' gi retest ii cqut'ncvand al the' gretttest cone't'tnratil ins i'l'ahle :1. lttuttritt tund -tI -DOt) were detected in 2 of 10 sanupleszuiaisved, bath ft 'nt Aetchergff sh. l2xpn_suie 10 Partspe'r-i.tihhiun e oimmiiiraiions of toxic chiot-Ittated pesticides similar iuu tb -,': ic tried hi the u''arv/ors.'vle ussue Irma ci site eul the fish ztijal'ed have been slinsen ic ca-use adverse eliu'et's in ratly lilt' stop'. tIs1 159ill).') huts, stone potential I' ur tl.uxic effects''! ,ualernttllv trttmtsiened chulirinated pesticides was pmivtble, although tutu-re c'lornstve sttldV wttuld be needed to tl,'tt'rimtine how probable such t'itemt arc, Ctuucente,ulii,,i a of in,tlerniliy traemshc'rred ['tIlls iktccti'd in uruithuern luike'ittimuuii1' tc,tr''.00l,'"te samples wt'rt' Just altirniing. .1 tile! il ccviii ditls'rt'iit t'Cfl cuangeners wt-'re ileincted lit 'lilt' Ii ittttm .1 the ututtpk's i'l'tthle It. t'cbt 133 scos di't:ctt'Lt tire ni.'st lr-eitueltli' u/tIl stinpiesu anti at ti-ic grr..ttestc'ncc.lttrzstivats. uup toll 2tsg/g;'t'alhe .5, POts 00, liii. attd 1:111 sect-c mcli deter-ted 1mm it single Ncwberg tIshi, 143 All P(J3 C Itiflc1s detected crc mono- or rfirtim- eongntneus ttungei loin I! .5 INSt Rut u to till IPUII lit.. In suhsLituied.]iic.c .oiigC?fls'rS tciicl to tie I tichic, inNIc than utiortho Imat PURs , 02. i!sfi. 0 pat icuhtr, ha e 2003, otul;- 2 elope-nets. Ii TI) 11.11 arid itS! liii were detectcd pc:n 101 was braid itt tin I T,:rviihs siittpie and two Xeherg samples at eniutcentrath 515 ruinging fri un (1.3! ti: 1 .2 ngtg. that coaxpo It C Li) the chit! rele L1lIilI.ilitC mi to- iid dii. irtho PCB congetlets 111,0 reduce the vomit uplak. the nun tOxjc isoitho pianutr lUlls Mean c1ncsntratuoits 01 PCBs ft and ILL wre greater iii iili ciilecteul fr in Ciitvallic Samples Wete lint Cit Ilecteil at identical cot inns mcli re-it to it Vt-IS lit pissihie Ii thin-rut i un wheu her difleretices In the nongetters dc tectenl is etc the i n-tilt if spill liti iii tenuporal duffetences. liy i all. ne 1 esults dirt Oil provide ctutnpeilitig support fi-ir 11w bv thets thaiclili 5 liii- .1 pesttn-i nbc residues or PCI! piosetul insunilcial sill iflettts ni-ic alikclv cantse for the grill ci skeletal defcumnitty lead itt eewherg 11th. Skeleimul I)itii iiiRle IIioassuiy I: lOver Waler Extracts. been shown t be less s(ritice Li the tiliflri-crthi ii:its than ntamnii k .r hlrct i..t I. Ii lOts evrn beeii ciiggcswd than from Ncwkrg luhic :t, (:ttver'i tic maul cliicnrnration .1 LULl I LII iiVuLrV/(.lic1e tLswie Was gimlet in Nesehsrgiisii Liable ti. liowov-r, teitherofihiscdiLicuritcn Wils stgnitiraiut. t)L) Lb basis 01 the samples an.iivzed. niaternal transit if Ptt- di es no! ppear to pose ii high ri.k of ovn'rl early uk stage toxKitv In \Vullamctio Rh em northern pilwtuttiuiiue;. PCDDs and PUflis wile detortabie in all ovarvu tom! samples. Specific cuinginers viricd -uinsiderabiv ining samples: then fore. a iiiiic C& uiikiiis zLpptslach !i used to facilitate atitibsis if tiLe resu Is md coniporis iii anacangsltesTuai2.3,7ut-TUL.)D IquivIulunis in &ICVICIIVOLV Ususun ofWilmatncttt River nuurtliern p keminnitw ringed [rim Odif Lu 206 pg/g wet wI Th greatest ItQ colt. titrate JI 12.06 pgfg wit wit isis detected to a fish front It lllst PCI-I ItO it i p p wi s detected no siitgin- Os ii lii t.ilL1ii) Results of lahtuatorv exposure of fathead minnows to \\illatnette Ititer tealir extracts freuni d 2 1 il 6 post-hitilcit v ith suhseqtient prune_rut to ii 213-30 post-hatch did not p r;vtie evidetie' that utnkiaunna contpciunds it interactions lttwcen rio t it L alt present in th p report-il extritcis ware ltkeiy caitses til grtaler drifornilve lead:; .iluseivct-3 lit lish horn rEam regi nt. ut the Willutniette thvir, Sliriviil postit. itch tanged Iron (13 tnt 100% iti all trials and there lucre no stgttiiicalii dtllcnne:es anmuingtrn-nutnacnts p. 0.2112li.73-Li inutiealillgih;tt exiractss ri. uOit aruteit I-x.ictsiarvaifutthead tntttiln's. SUtilVill nlcititip Pt- ce-rut was variable among luaostvet. tiiiiitarmiiiQ roncentnatiitisweicnearlv identical brush collected friumihe two stud" ties It.) itt r)21 Icrils nepi mutes aiiul aliting in alt.. t alging fri ni 5 to 113 IF-h per replicate tI T--i3;,i - itt till COsOS, ii LiLt ii ittim of 20 fish per {L85f.0.31 pgfgwetwtfturNewherguindcoivaulis,t lable :o Total concenirall ens ofT I :Qs wet e less that di expectnilcijuse Li xicitvdurlttgearlv liE snugedevel ipttw iii. Oil the basis ifjne,istiitd iiI)i) crmuceiiti,iuh its in fisii eggs. treitittient pit-nip s-crc examined for deformIties, it wits not pisslbie 1.1 detentniiue whether fIsh that cIted during grutwriot Crete defortuied. Nonetheless, the itick if a ugmuficant truttui-it-ri'l;uted effect I in sttrvivuti id tie ettiliLiliaiitl day iuilih suggests tltttt mIte inirmalit tliiriuig giveLII tilt. raitdu,muui; ilistrilt.lut'l lolling ieplin-ttcs all diul not the lowest hserred it-fled connetitratiuti tot sewn itifferen fish sJscies ranged t tan 271)1021 ) pg. gaset vet tu XCI were - 175 iag(gwei vet teSt. The3 EQ coneentiatlrtns deteeted in this studi-were at lest 135 tunes I ret than the LIEU lulv the 111051 sen sOlve if ilit sewn species tested li'3. I-n thin morta.thcThocnn ntrettinsiteiceted ore ,itleast25 Omits less than the prohribk iou-i iiiserva Ii lit-oil eruut-effeu t level INOARI3 ofTt-Xs fir lake In nit. irlitch is nudely ic-parried as the flush species most sensitive lii ditirin tittil dioxin-like toxicity (CS CS! Thus ci.unceritrtiti nt. if nut ttriiallv ironsferreni PUI3Di, mind PCI *s dii not appian hI-tic hi eatisi rarl life stage mortality obscure -a tremiunent eIfe'cL. When stOIc Ic- drirstiH Colt iruii cttrv,l(Urc CCitt. ineludittil as a deli irini IC.......25'' ot the- Lislt etut nliLled were citissitted utt deluirtard - aitiiuutglt Ill ti -urtetlt-llependt'tut ellect teas ti:) oItsenvcul i. tiii'iui \Vlwti liii tUitliV'ils isis restricted to nih- tb -se stelnritliiies -lior,s-leitaud as qii;titlativeit' slmiiar ii' thrist----F-served itt Wiltutittette River htsh ;tts pr I ca-tegoilus uiefimmed F- i tIutningliant et ti. 2001 I Xiii, the it [ruin i.1.i3';-2t for the entire itit-irlence of de-toritti ties pcupulattott survered iii c-.inI tuuai, Civen Ic_still sample st'es On the basis cut the llter.tuurit, it tout. uitrletur wile Uter attv of the couwlittationo --f Pt )Ps ileite ted in VLiitaiimette Itiver nif 210-3137 fish pit inlal. ils represt'nteui 2-8 fish, In ulli nartiicnipikentiiimii iiivaruiiiivie tissue wuihIle likely ii n-I as-si vial! ii witlt lily psttictil.ir treattlinuul suit esitient_ The u1istnihttte ti ii dctvlu:ipiticitlol seuirrwls titlaffected catusi'skniettl ii [rurittttii's. it vet-sir. miiisigtliIieiutt ii liii utters lnmaieinailvtiuutsfenred POPeiitsemtit.0 otis usi:te.ibserse.l rases, deformities inn-re sprr.tci ulern:ss ireaittieLtts, such that Even 11 early life stage eapistite I 11 tlsveis cattt.i tip soiiue mi55O,t3tij httm-t.ituuentttttiin-NI0(T'attul.tC,ttLoi'-. with titost fish ltadtig detelipitteittil u ures pleater tItanS. in them ml ;\tikd. fidu trim tile Itt :giuuip terre slgiutficintlt' disruption OfCiUi devcluipiiietit, kihimtg Lit skeleiai ulrturinu- moie tieveliped 111am) those Inont all u titer rreatnlellt greuups for fish front the Newberg veisits Ctea!Iis sites il,ilk lies, ii was unlikely to leciltit on 3--.-----ilil gr-ilct roies of skeletal deforinitit .-\-, a. v-hole thin-s.: ut-nut. iu tided nut mtnplhiig eviditce Li sitppinni tite- hpiititcsis lit at greater maternal truinsler ut It Ii 5 0 ,as.i Ii kiilv e.ttise ii the gre .i ten incidcnce of skcletii dcftrnlitii- III tishi irttutl die \eueiietg region of the Wiliancnitte River Sedluwut POPCAsmailuutniher CL Spot Site per ear) of suiThdal sediment satuupies nuetliecteiti finn Newberg and Corvallis tites were analyzed for persistent chlorinated pesticide reshities wiui it ;Us to deuermutie whit iii tropism transfer of these comp.iunds 0 in sediment, or direct exposure of vnibrii-lauu I tish i tauitculativ fin lii outcast uspawnirs; ! tOil could ItcelItIt fur tliifereutces lit detiurutittes at the iwo sites. Chili initiated pesti ewtnt. suet 1101 detected itt sanipies collected firm i ucitullis on \ewhcig itt either au 2 nr2uti3. 1ii2002 Putt-ti wasdetucted iii a;:I t:ireailissanipies and t Ut Newberg tuiuuiple. t%.iticeittrali,ns it too it i-it-irul ijuan 13 LI) 66 ng/g Addtttitna.11v. PCI! i2tlaas iliieiicut iii a single Corvallis sample and PUtt. Lit. 101 . utuid I hi:; Wilt detected in a single Newbergsunpie, Conceiiiraui ui if tiusi 0,02-li, tI,------en, Ill cttiiceiitratiimi-ulepeitulenre nuts evident. As a whi ii-, there was mit t'tl deuce that Willainrtie i1 hirer Isuiter cxl racts lilduiceul skt'ietah duefc.tiiti U-stir odterselse adversely Iliiietu'nt latvil lillietId mttittncuws t'N)uuistll fur Itt! h from d 2 Ii, di! lli)st_li.uln 1, It flutist be it uuteul, hu;u ever, that a tlC:Sittii-e I espotise In the skeietil delontuties Iuiuuassuiv lid tilt rule out thit pit.sibilitv limit chcitiicals ctjtttai.iied tnt the exinlcts had ihe pitritlil tiilinlildl' ulehirtittlicttncvpruttiii tiiit.Asdt'sigtted, the assay itis stile tm pm uside ut I (its ti able cci ct-ti fir the p itritim,iI it the riser iu'aiet exliaei ti disrupt ii ow early lifesttugl' ttevr-l'ptnnitill pt-ici't.sl't. iittuiirtatlt lit lurin,tihuil of the u'ssllii iii cituhual cilutnti tin assay Slims lltit uOipn'cted to be iii elleet lit' sun cr11 fur cllni mcii'. tide tn ctmlise sI-el etai U. fuirini mies through acute- tlellrtinsttscui;lr damage, A timesin s-s Ii ir skelu iii tlu put i'nl in Iii \t lucid unnlei ass a" umoliti itS sh,wttl that ,is ,-arl it, ii 5 Ilost-hatch, nearly all fish litid ossified sI.ulk-,ittd Initial teridural fonttialI-.ni, as lnthcttte-d b tassilyttt run it iOn aultenzr)r-n)eist centra fun- published resuliti .Atteutpts to validate the assay using Cd. or, is, NO. 10. Loom Et'ivIRONMEtIT.SLOCIENCE &TECHNOLOCY. 3503 144 So, and ehloipyrifns as positive controls s'cre unoiecessfuL as dckrinitiL's ro not Induced atnonlisle )nicIltt011005 hmlltbhslwd results). Robust application ol hi mviii Idseill require additional characterization cit th LIcILcILIbLO niLelianhsJn cit acron and further oplimizition to ridmin' Irtality-related vain h div during grow-out. Ac,cIatia oi ParNtes with I iIinhitie'. in kid QIIiX.(d lisli, The imcclivflrv f keleto1 d1-lonuiliis In Wllarnelte urn fish was siroiigl lii k1'd with uci.occl cailac of a digonetin treuiatnde, likely oidiflii.s d iii0i (317, 17 4Anirnalvsisufciriirrd end smaincd S1L5lillhmlsrf ii' 'nhins ptkemlnntw; and rlttii'lni nOli c,illcclvd trio (Lii Wilbinielti River locattoits. including \ vilmeig. Wtiwitkmi'J kily and (,orvaflls [7gnre I. rr,mjcltisjed that Ilic' probahilimv .1 having a precaudal skeletal drkrmfty was strongly dependent on the number nt trercatodi cySts in ihe lush 'p mIs iii and the location in tlm river Ii 0 (100 01. ipecirL. and fib size weremit sigltllicani pri'dmctoi s I 111u1 I nn,iti.des were dirctiv assiclaLed vith tilS.5. iiil2 pFirll.ur picull irtuimmuts dCll.LLted iii ehlscliu,iith and t7.:C inortimeri pileinintiw (it 323. Add.IumnalI\ a 1 Ii viohis sp., Orb cvp,iei was issu.iited with a sigiticani pvcrc:nt 7.3'.. northern pikenini iv wHim Insiilgicil!v snrifi,ihle skejetil de1urmitie- I Ui. 'I1IL' rr.ul sIIltge'tvd hit sirisiles i;r a likely raise tIlt the kihtil tifnrnmitivs 7 -rivi:d in Wihlantertim Ithier lish, I luu'i'snr, sick on tile bask .l exanainati on iulield cuillectod specimens Ii was lilt jLIlieitLlQ to doirrinine whether piOlsi In's wele acnmalb using ii- formlitrs ui whither dciujrrn1d fish worn' P uplv In J vuinciabir' LI infection skeletal I eio t mU k's [lii iIa V Ii I xpi 'n i' Ii. ti;diui[Ilis ilsnteitsi eIarilc.Itssullsftlinl,IlL,,It,urinfeutI,umstIIIILs cins-incinglv dentuitstr,ited unit Sri tsltt,ii ul,'Iirnmttin'. riOmiiitnt ivith uSC .,IssI'VL'I Villani ,ttr RIver fish ru Id lw caused tiv tieuial 'ic ceLLular idinithecl is 'L0;,5i:s i,ea-mia (Figure 2i. F7vi' separate exposure trIals u' conducted wS!i t1iilitd intilniws 1 L'vpililid pL:CII.S) rulgig from ti l 23 days aid psi-hatch' Mortality was variable and Uea hIgh Lit hu II cereriar-tapsed i-I 17(.i art control fish 5-ill. \i.,iinlieiu'ss. ci.,ttchisI a', c.i&ild be drmrwn,A high i iteidenc,' of iitlectii II 13111- lOUt] v;is hserved in coreariie-cxpusrd tilt front all titus ,,uid tnft'cieui lisli exhibimid a high invldei:ce 'ui veitebi it itehritmiucs ,Tmi- Table 0. 'dust deIitiiltts weri' directly ass' ciated uc!l,h meracercaijac' utigurs' oIiul many ill trenillodes wnri sure not found in the viscera. Si iii hr II) I(IO[?ihJii:l] iiv in the pi eseul studs, .3 L,,'ds in 'el low perch .ipprcunthv LIII'S not infect the visLim al rgoi sOT!, I ad n clii. I (3/u ,hiserit'ed bony osm,lcles Ins-chow perch C'd hr,. iuim'd. Infections brother nietacerca.rtae types Ii' n brett linki,:d to vcnn'brai an nina lies, ytmm'cle tnic'ctions hi' 'bu,'i(l as nod ,io 'ninu,s caused vertebral defoitnt ties In cvprinld fishes ,iIuy , and Fiilwcioce sp was suspected to be a niiilir cause it slipernuinerarv linibis and othet vertebral chaisgos seen in Norih Anhn'rican frogs (Ti]!, Thus, ta"tli empirical e'ddence and literature rt'imrts simpprurt lime i'neiiiion thai tremarode parasi Is are u'ulusimig skull-Id huh 'niiiitie's In \Vlhlamette Rixvr [LOt, 1"iittn s' linesuigauin. dud ugh pa n-intcinleclionisltkvl'' the pnmm-iimts' if skeletal defo rttiities ii \ulhtnmn'tte Ether lishes, qucsti'Iils ri'ittattt as i tehetlie'r sp.ivai dttln'riiievs Lire due to itaicril or illlolpugeimI f,ictoi ' icremsed nL'curti'imL'c ii mwinatide itivc'livmms hive been iiiim'.h to rmnmrlui:pgcnic huvllnttiiu and physical ahmerati,in ill aq SUe Ital'itmis C1isn'li hL'u'litlin.iIl acliu'itIm's i..'. I'iIeIttiihsIin'igisn1 lmr'tti.,i'it i'spuismii lti'rbi.'idn's amid pesticides .ini susurepmm Lmhihiii' I in 'gs I niiecumn hs'ilmctulcu'run.inw it li!m't'iul n.' unit l'i. ': 1, 'i. huts',' il', hen rep 'nird i 7,' N' imie 'ii the chemical cnlutuiitant, dL'Imci':,( ii this study an' kuwn to cittsc ilitmnune sllppressiit ii linreloe SIISL'i'lttihihttu to iuficiiu lit at ilin e,unc,'iutlitl&tis uhsnr'.'m'tl. I lluwuvii , din lii 'ligI cal assays used in this sttiuli'dl ii, it test the interaction hut Wi''Il c'xp.silnv is Parities amid e'sprsilrs' lu' cmmtiphex iatixtilil's ml 'li'inii.mls pit'seiit in t5'thl,itil'ti,' ttivs'r w,itn'r svdilnvmtm t'atcicls. 'this would h a usiutul step lultuid dr'tu'rnit iii ig it'ht m'iltcr tliu''.e ehnnit cal'. pr till 'lii U'.ci'li Li iLhIL toplr.isn's.'\lin'lnCmltLei. uitsp'ii'.stlti that '.patialdihlu'tint't's in detdritutlu's rellcu't Li tiattiral Illicit tfln'tlOti, th','1'ii the life 00, an:,' hiimiOt charimctens ics cle ol,tii,ims'ihia las's lug tithe time iimtu'i inL'didtu It' st I snils such is / i:ii 'd IT lhedc'liiiitive lust hsh-uultttgbmrds rould rL'sti hit gnu'uie r In mini i,.if . ahu lid iunce mind p utenU alt'' nile inlectimis N mtilriillilctuuns, iiiilau'ttrimmgllin vi.mbiltivatt nwuhens of ttii':uuhi,ii lid iulilw Ott Its lgm'ltIs stielt as (ill]', nun- also plavar' kAulditiuiiLIt 'nil' parasite ecohugy ui,i potential int,'raeti,Ils suith .inthrupogc'nic' ititluences con Id Itch dr'tu'ritalne appropriate nianagentetat actions br al l::ted rngiinms cif the 3''ilhanleitl' lOver ilIStIl. .i dirL'etiv located along dii.....rit'hiral column, The types ii deihrtnith's observed u'eie ahsi.t identical to thse i.ibsi' I'Ll in fteid-colk'ctc'd sprcinieits 110, Ji Including 'itta sl'' ltsrdois, fused reticbruie, anti itereasid vertebral deiisitv (Itigure 21.,"ts in hrld-Luilleele,l spectnieris. lumu'taeu'reanam occurred direrlir appressed in or dt's'1 i';it liii vertebrae ,ntil were often assnd aU'd with hi ilL ll''pertt03ilt\' lit cmntttst 1.1 eercarlac'-eapisedfmsh. unIv It if tilt' L'IIlIIuI ic-h CXiliilleih exhibited skeletal dt'lornmitles Table 1!. CoWl ii delmimi tins were characterized as riIfVatutli' i clii' spine fused vertebrae. The Incidence o 'duetn'tal deluintitles in LI trol fish was similar to biiekgj aid rates of skelebil ulc'lLiruulies determined for lab-wind latlwad minnows ,x,iiitin,d h' ilLiclrericenee micr.scipe a'. part of cxposules I. river water extracts. Coutrolled lab at nv vp sure to .3 mInOr: LvplirilCd vcrtebraldoformltiu's' lusnn'e,l iii fish eohle,'tiuliriit lIme lild andfiirlherdemottsmritid Eliot this pirasitcwashikei a nlIJr cause ol dn'Iortnitles H 'i:'Illwiie.tte lIlvor cvpriiid lilt. usfs hcterph'eid digencan irvin. aIde exhibit'. broad hi spedficitv. illuu'ting mail s3i'cIes in liii Iainih 1' priniiac' Lu'. as jOb tint sc'vii,l then tnnili,s I/Ui As iibscr','eLl in both our labora tory and field studies. the parasite exhibits remarkable affinity for bonn' I Ui ITu. Most of the meiuic&'rcariac were associated directly wills skeletal structures and 3504 LISONMENTAL CC!ENCE & TECHNOLOGY 'DL. 35, NO, 10, 2t05 ckn oJle1Igui1eHhS this c ni wi's sttp[ rind i-v a groat loam the i uregmi i \Vaiu'rshn'd I mihaitc'eittiutmt li ard (ni 2uit -502, ,.\dlhiti,inal suppmi'l. irs tim,' aspects nI the stud\' i 301' tdemi liv Nit] IS L7_'ntcrGrint l,Tiu I:Si3l:Ulitlirotigimfliegnn St!!mi tJniii'rShy's Matins' U tr.'situai,'r ttiiiunc'dictt I titer al hr diii Orngotl Agticultiir.ti Fxperhittetit SiLitloil. specimmrns were collected tinder Oregon sm,'ieiifihit takitig pvruliis I Itt 2002-1 15 anti I 13703-117(3 amid aNu iA,\ Fisheries 1 d authorizationletter, 'V also thank Mark \Vilh,tIns amid l)rk' Johnson. Supporting Inlurination Available tm List cii all target attal is and p.iratitciers analyzed Liii part itlnsirimmititining with .1 Vu/I ('Pill Smile umoIc andimi situ stmnphng et hioa'railahhu' uiiinin' compmunmts nd metals mmsimlgt"st)s mild 110 F'.. 0,0 uhiitnmimilf I] ,!-Lahle-cigc-n'Lihlc'arichot SL'tlip I:; 20112 and 2m,um:b p11 id uiIlt irentis. IIm list of target ,malti:s aimaht'aod its riaryfoo'1u' tisstmi's arid sutlaclmI sc'dinie'nts mud 11w' eindiittr'ations duin'ried in ccli uatsmplc. iS ck'v.:'lcupnietital sconilmg ci itenii used or t'athiead tastnn(mw skeletal dcfnuttties IS5LtV, and 61 ('iLitlTphes II dclrrnmmties nitseryid tnt fatiaead tnttltluwskek'tLIl del irtitines dSSOV 't hats inateilal Is available liii' of diargi'uim the Internet at htIp (pubs aCs 0mg 145 clii t'ut(b'iurit-p.diutxitt tunxiriivin the rc'brafish enttbrvte altered uegtruuul hi-id f1tiu atid iutp'aired bear (.uee du-wlept'uneut. Litetature Cited Urkh, l. A.; Wenti U. F, Biirennieiital wiling ri the ftTflIantitte basin, Oregon. WriTer Riecureri Iii nnitgaTiorto Ileport 97-4(123. U.S. GeohgKaI Survey; Portland, OR 1(1411. Wentr. U. 5; Bonn. B. A; urpenuer, K. Ti; BunkO. S. H. Janet. 1.utenen,.k; Bencala, M.L;RineJla,lr,A,;iJhrirIt. MA. Walte,1. K. H. Walex (1uaiiT\' in the Wi ('uiuuelte (aint Oregolu 11(1)1 1(1113, C.urcular ITRI. U.S. ('i'Ogui'aJ Survey; Pordaud, Oil 15)8. H.; F uaeui.C. St.; \Vuul.e.I,tT. S of rOut on Ott aquatic biotu and their I ibilat' in the V'lluiutiette bnaiu, PT) Altiti Oregon. through 1(115, Waler. 8cc Litres Iuvesti(1atioos Report 97.4033 U.S. rr.'lgiral SUt'VeV. Pihtxl, OR, 1(1(17. 14) Ilenglnetcui, B. F. Stof girl I aliLittles. ci Iucclallv skehetil deferniitieaiti our tush far ni 'tti triItg it run I1l1uitiott. Phi/on. Truute, A Soc. 1.ojok, _tJ 9711. 2Sf;, ii? (5) Valentine, U. IL.; S iii,. ST..iiut ii lIner. P......ninetry aunulvaun itufishes; rt1ieu.ublrsl,ttusrk'al ituulii';i leiinrottmetttalatr''ss. flJt Ruth \ iii (lIT Ln'nily.A, r3..Aiet'ai.fn:lll(.:.vrnd..o lol'(".'.ilui:ititl(1ilt)ntc ofieleuntiuu out fish pcnpuulnui;' L,to.v2ti. F in.....' 37. 2(15-21181. ulics in fish fr ii f3llutad urfiurewuters. Aqtaail ToxirnL I51 21. 57 tnoTcu:uli,' baue,f ruittitiuuna-itttltured u'li'lu Iitiiii'c lit zu'htraflsh c,ntbrivuo. LoCrrar Thu/wi, Ot,',it. 211(51, 19,:tb'tli '.1)3 I 2(1)1, 261 lleauuuiult, H. ). Lethal p1'1 fr Ihe wlti 5' si.n.f.t:r is;'uttus,'tntnue 1172, liSt, 'OCt51 cr10/il) iscapedl. . 5; ia .4ieu....'; (macside, H. T.Sciuie t-th'u:ts cii rcvgt'ul in nu-luihotu In' tetitperatLuTe 'f lkt' trait titbu'u' 5. (il/I /eui/. 1)51), .57, 11ii1iS0. Abdu'rdice, I). I . IVickeit. VP 11; Brett, 1. It. Seine effects of ott des-elupotett I 1211) ieuttjennarv .au(u' cuuuu' ti Icier dissolved oxygent levels en Pacifiur su)na',tu eggs. / !'utb. II's, Staaa'd Coat. 19511. .15, 229-2411. Cahtriel, St. L. t',t,'iuts .uffa'titugtlta'niti'nber and Bnit iii vertebrae i301 itt .Futnduti:ta );u1t't'ia,' 11/5, 1. t'.' Zn .I '1.14, .1, 1113- t33 Ks,,aiti, IL'. F.tnbr'uuc ik'us'llnuuetil, u';irh'(1ross.ih,utuuul ittenistir II) Tibia, until an investigel kit if pliulbIe unities. I P08 .ihIu/. 73 /17) Muore.C. 1.; liBscuut, LB. luucideitt'e 'if rab (venl.Itril cctlnnuzis itt adult Pitnt'um tiiverwhitc'perr.h, Mntnaute-Atni'nicana. Pej.'cB 1977,2, i.l3S7. r iuibu'a intuIt SOlute gutitcifii'3 exposed to rtauuthiuu;ti, 'nc) light uuutettsitv and trntperntulre. 1. IS/i. .Rn. variatiott in I (B) Bautnaut, P. C.; Bane tent. S. J. Sen uhral abnornirili lies iii white crappies. P noun ,'tetaiiti;i 1.liPc''itj:ie. froze (the Deuratir clietug.5 lb.; UsiA. If. ITs So, Cli.; Wit, Ii. S. S. cellular and 25) I'to(itar, 1. 11. tig ;uitatl Iat'valsutreivuu) ofwltilu'stt'kt"us iCun.otrIeLtus contaucraouri at liter pIT. I. F/alt. Rca. Suxirn/ Luau, 11177, .34, 2k2- I I (7i S1oot VP Skelu'tuI nut Tta'i'uu/. 'i'm '2111)1, IrS, 1112....150. 12-1 33tttri( I -'s. 11177.....- ;'.'t7 402. Si) FlaIr-n.). I.....-5.lf.':. L. St.; Snnith,t(. i),3scitrbic add requirentenis 1 nr.hui e.tluu. 'I .inI t;uuttbiwlrctut_ Traut.u.,-t,ei. F/sb. Soc. 196(1, P21 Kl'pl;l'l, '8. \l.;Post, C;. l'Tislologic-al ali'rstiotts itt trvprnpltait_ d','In-i.;'rt ra;ubesc' trout, /, ,SSulr. 1075. Ic'S, Ii;)" lletlgtsciuti, A. Effe.cisof bleanlucol pulp null efu'luettt sti vertebral clnferla In touritoru 'tulpiu 3! nruru)liiul!:us 1.iiii1r.vt'O1s t.5 hi thi CLIII of Rotluuia,Ardu. F!wlr,.11iioj. 1.11, 121. '3 'at. 15':; Stt'Kn'. C H,; ClIerde. 11. Gene-tie vari;u I'ti C,..,t ci,o nil di'fc't'iuity I.lndnuifd, 5.; Titutlitu. I. A skeletal deknr.tiice if r triln'rit pikes (Eno.r (urine re!aued to a:' lutz)) ai;s. 0',;. 1. 1 aIr .4u ui;. 34) P;tuivton, S. IT. Venir-brail colamtt tbnttnttnalili,''. itt brown Irolti, Sruhitio tr;ttnt t.. 1. 1';.slt (lie. 1957, 1(1. 33--ST. 125) Priori, 5.0, 111,ouiiauueoiisskuletaldniionttuitirisitu thezebradank' li?ra.:i: 'Stir's., icr/i tic-..) fi.tr fish tcunicitv test. I, 1-SOt, B/of ISIS, 13. 71 ;;3. 361 Kutul.M. 1,.;) a..ft 1.81.; St'uicu;n, I, K,; Yasutake.W, 3'.; tOlL A. Spinal as-.; In in lug ttelt;ivt.ai titue to cranial otid erriebtul lesions niasectateul t'.'i: It C.'e'Ol)ileiiB/ 1.'o;','l.'eu1tb//a infectious in sahiticinid fishes. D.0......('alt! (5,5 hIltS 15. 113-121, 137) L-ipatra.S.ti.;BsLa. 85. N,; ItIL'rlia'l. K.;J.aiies.t: H.;Shiewtnaker, IAS 17.; Wiittott. 1. Ii. Nu'g)igihtli' risk ssse'iatc'd Is1 ththenioveiuent of prude-ic I in i;:t',us' ti' ut, (.a.""'tlIi't.! 'ii tue uri'kiss lWalbtutuni .ca. is'pi. 4s. in 112) Ellis. S.C. 1. uI wutrul,' .qvt'st0'L/ihttien h,' Ilit,sn'llnao. 1','j.o"t 1.rti;,'l )nt),ol' fa';niflit'fi( I. I''' t15.t1511' ,'(iluI;i,. l., trt 's l:et.'u.' No. 2/53(1-03. P1'b, ti, 2ftiitt. lithe. lWWiC.dt').5uiite. 51!latttenti'.seqsl itdiei.e.11lrn.hskeldetonunrpt .11. (.131 Elba, S. C.; Deshlet, 5, I,; SEller, 1), C1aract,'nuaiuu 'Oh aowrub]ages Jut the Wlilnuitel.te River. Orap ii. toittg three dilfnre'ttt. bie;tsssuatent techniques Ta trh'rt,:u/uv!';'ut until .Pe'ausrilS;; Laeute.n, A., t'luututl-ite. Iii. l.;ds.; lewis Pubhishets: New \urk, NV; 1117, (.141 Mnrkhe, ISP.; Prettcli. IL; Nujittan. s t:lcttuItutl, I.; Batik-Id, M. Hislorkal piltet'tts cii okek'ta) let 'unit ti's ut 1 ii Is's firant the WilIatnetfe Sisal', On'p ci. .Varu)uuu-N'r,: Vu:. 21)02, (13, - 14. (15) I'Ioiconube, C. 88'.; FouuiL, P. A; Leout;trd. tiN.; Skkiun, J. .81, Iaung-tertnuinflus't ..iif Laid ap cure out i Itru' gi'tierati ttsolhtrontk trout fSalu'eliuttri totiitii.ul.,i. I. 1a)u 11,1 ficenu! (tilt, 19711, 33, 1054- IC/ill. Ilk) Hudson, P. 8'.; liii. I, 5''.; Blunt, 11 1).; SInger. S. 1. lill'erus iii dietary aserrb 'a,' it itt chronic lean t.cxu:iuv IcyliutIg tuatbuw trout lSuino 5i1rol'ui'. OW. j. cnlu n rut c '- 1 uttIL, 37, J7) .175. in .8 iii ii,' sal mciii, .Suulutus ,calu u; .tij;5 o':u/:iuuu' I 51143, .30, 210- nnI'ecui.,iis h'sL'iti;uta1tr,i'IiC ti-ntiS virus )tl'INV) ettlenni,: ,'t cc's. I, 1./sit. Dj, 2001, 2-1.351(1 .117,1, FIuil;'irk. it. I".; Eb.Staibonilt,M,;Adkusoit, SI. ."s.;Ma'i.ontiell,E. Irtiuti atu 13(1) SVluirlL rtg di cL'.isi ru-cltt ergu'nce ntnicutigwilti I 'itt. I;,'n;tieto/, Re,, II-),. 33 '.17'' 139) ,S'lntlI,''uni, I. 1..; itt-vu. A St. 8'.. 1.arts'eti, F,'.; t)ishcja'htess'aru. K.; Rr;'rs_ 1'.; keot. 33 I.. P'iu'n,'l,'llt.ilttL un'ilm1s:;;al lug., tt.ip a ttew gu';;iic i;iI sp','i'c'.I' .'sli'nc).'ei'itll;l Ii'ouiu Itt' u','iiiral nn'tan,uc cycle ut of the zebn,ul'i'uh -:. (('.1 n;COI. 8. f.ui i,.\Iirnotuiol, 20111, 4(1, 23:125.5. il/a Cntuuuiugli;iuit, 33. F.,; Mat'kle, H. P.; WaIral, V. Cs; SB/itt, 33. 0; (17) Benglastait. 13. H.; Orion. A S rrteltr,uI ,tel't'uuulla:'s id pI-slolegiculi-t'fs'tsitu i'iiit'Ito"ls'nlpun uSt'eptt.ituusqttudriu'n.:ruilsJ .inuuu)aieul Itc.iu's-ttte it) - couztatiittg alterhoutg.l into e'sposuur' 'i3 Veri,'lir.tI l;uuitre3' to Itlitugills Lii 51cC_tint (A. lisper tO.. exponed to usrnii civ t ruin luui'ku.f u-sl:rilec. 1' iu Its, Ii ii, T''.It. Soc. 101: .3t7-322 itilerit. .'tquuuui. Iuyfrit/ 19111;. Ii. 215 I (TO) McCattn. l.A.; Jasper, iT. 1.. \'u'rirh,r;I ,Ii'.uu;uc,'. Ic bluegill,; espr.sed to acutely toxic level. cf lut'sfi'Id'i. ;rr,,t, Sit, ls.u;i. L's'. 1:172, lOP 317-322, (15) Heou7) IT. IL.; Spitsbergeit. List.; tIn uatg. 53, 3!, ;.,kbit't, u:. C.; Perercon, tI. 11. Furl)' life siege toii'ity if 2,3,7,8-I..,irai:Itkiro- Curtis, t., B, Patterns of fish duforntltles and their association ertult ireiuunic de cysts in ihu' SSllla tuietti' River, 0; ac;' 51, !Stt,;lruei. 3/ni, Re'lien, 1413 I'huckles, I. N.;'t'uthnrgu'ut. St.VP; fd;tntuuu"yi';t, 5. P k"nuipu-rttieablente'iabrnute dcisicu'c,'t.ititaittiit(1 ttti'.tl'l luTil 1 iirov;ub'tur'.nnh to ttiouinaaniugtlte biu.ivLil,ubilit\'ot'ltp,upliilii'u'olti;ittun.titIs ted estiuuiuuu:gilau'inlui'e'o ttc1'uulraiiunupot'tll LuL 1 l:tuiqtb'nt' 19013, 211 523-552, 1.12) dibenzo_p.dinxin in ec'brtliali '15. tn 'lu/n. 1997. 1 Mehrle, P. M.; Haiutes. 'r. A; I I.nnili ut. S. I.; I.uul Ta'. 1. 1..: Slaver, F, L.; Ribie-k. M. A. Helrtlotishtip bu.'twi'i' a hrdv tiLiinunatils and bone developnueuut in eaat.cc,iul sit 1 Itell line. hiatt. Aor FisT,, Soc. 19182, 111,251- 241. Mayer, P. 1,.; Meltrle, P. 1,3.; (maTcher, P. t.. Iu;leuactiniis of t;oxaphe.ne and vitatetun 1: lit chaitutel caiilsh. Titus. Aunt. Phil. Soc. 1975, 107,32(1 33:3, 122) Olseon, P. H,; Westerlitud. H.; TeL. S I.; ttihlssotn, K.; Berg. A. It.; Tvsklind,, M.; Nilsson. I,; F:rik',ts. H. 0.; llitttr,rt, U. F., liBerIa ofiouik'nial exposure to estrogen id rs:tt rid ifl'erertt hifestages ofzebrafish (Daub rCrau1. .'ltutlt, 1115)9, 2,1, tOt 10(1. ii S ions nina S Okult its I (231 1 'rioku Ii tJ,u,. 'u5, 1) Nhiyama, iLl,; Uetio, N.; ttein'rsu.ttt. Ii.; Iturag'i. 'r. 2.3.7,8-Telra. i-83i It uck[tt-. I. X.; SIn tu uwto'ru. C. K,; Peter, 1. 0,; )s'Inck,'uca U.; lab', A. t.iptd -cr.itnt;iiitittg, senuipertnu'able itte.toultrrnr.e desires Ion inonttol lit et cii i itlantinant', In lIe-i 10 inca/i 'ti i "cleuol 1993,27. 2150... lIST.. Seibtajlt;Iaruu, I.).; ltllltcs.ilt, l..; Loper, B,; Anderson, IT. A. Ilivarnuu;r; Hui 'it ''-It 'ic-I chettitcal conuiatttlnuttto iniishufrotu th5' Ililhuitt,,tt,, ibsen. l',t'ti,izud I tin'S.;, (Sign. At'di, Eutuin'n, HaitI:. i,eu'o'uP 2l)'t. 'U-. 114''' 133, i4't) Zhu,It;g. 11.; '}i',')-iun,S',' Pu't'I',t'tttsn di' rhuiStdlu-nisticsnfulitfuisitun gradiciitts in tltitu fins t;r tlte li-situ tnuOacunu'tti('ilt cii Irace ntet,ils it: aqiter_luis siuiUtir_,it. A,si/. l.j;ult, I 5457, (7,339124117 (-13) tu'otpiui. 13. W.; Ftirlottg. E. IT.; Steve;. 33. T.; Tttutrnnstt, St. 17.: zaugg, S. L'S,; limber, I.. II,; Biixt'tt, II. 3 Pherttuau:euiticuhs, hcinnnotirs, and oilier organic Is'uuciu'water cclnutat'uiulnrito haLlS. slreattts, l99CI-'2810; a nattenal r--;'tunuuuluuauui'e. Stun/mci, Sr/. TnOt,te/, 2002, 3)1 1282 1211. VOL. 20. NO. 10. men: ENVIRSWMENTALOC1ENCE & TECHNOLOGY. 3590 146 (4f Do. 0.1.; iroirkol. V Kiudahul, C Zuhar, 1. iauuliarirrg iwrnral and dc'ior'iiv hera' cieveklpuieat In zelrr l3slr ernbrrs usirr (oUorc'so'iri 'Iii eeoc hare c cola, JAr.'. Si 2001. 236,239 246. (47) Koru, M. L..; Wanal, \".; Whippu. C.; assesoraerm Cim',rrnrrp.4;n'r,' 2(1113, 33 IFi, 1110020. l'pslfti5e l't;it'.rlraarr. hltp;iiwww.pesIir'iderufrranr6i 011 )'i.535 titdex.htrrri. urmingharn, 5'!. F; Critrienc. C. fl. I'leklr'(. I. F.; Cur0r, L (I.; Spiledic.rp6'n, 1.; Markle', Ft. A iligenerarr 11 iii ova nrarinmrjclrraum: jill early )jrrough juvirii1e life cvole tricercariue i.4.n'er,';ia '. 11121 Walker, 13. K.; Pcteis'"tr, F. F. Pateaic'rna f palyclrlaerrraled dibr'naeip.direein . niilrerrerfuurur. and lnupl;a;rl .:nugerii'rn, rm'l,n- rud a urywzaaa 13!orrrbohiasp.i rasor lirid iLlivrrrtebril ii i'rriiirie' in cvpriul.d ftshern frcnu the Will burette (hirer. Oreorl. j. 'bps. ii. 21K!.!, 15. 110.. 121. 'icr. (('a Ith Noah. LI, F: Mucy, 11, 511. 'tIm Ftc cycle arid riterielrv Ia muir oi.hpalrediria ne cilia (Skrjalrin arid !.irr.Ii..p. Ii) IE''ut'roirir. EE.nl3, 1'ln'tOrofil.,ldaci II (1I'ii9eilt. PT'rr'rI, if.'.c:;rl,'ae...... 14'ijrl, 1974.4!, 223 2211. (413) Dinp,erkrra. C.; IJirier. 1.. It. Errzi'rrrccIcarirr üfrli.ur biLls qiltmnri whale rrmaii r'erlelreara'. Cr danio.rnatrariarr rat carthirga. strain t'r 2.3,7,11-'. b)plme.rrr'I r'nmp'rrerrin.rsnvinlnrm earivitle 5109C mi,rorl'ilitv in iiilirbrtw trout '1 i,;'miri;e,,'r'inrsc nrikias).Arfninnt Tcnaiccni. 1993. 31. 315 01) tart 'deem limp. SI,: 0ii'jrbnemj'n, I..;' Bnjsveld, Jm,'l'.,' l3nmrralroru, Ii.; Ci',)m, P ; t:r.I.m' lnl.; 1'.inav. 1. 6; !'tarrberg, 1..; Hensirguwu. F '131) Potihoff, 1'. (lear rr Kerr rich', a)',',; )irnbtjk, r.; Larrunart, 1. C.: v'.nmr Len"uweim, F'. N I Lrerrr in K 5 11 ii ' ti It I P at limo r I S 1 S Scloreuk,, Ft.; '1111119, Ft.; ')'rskiind. St.; YrittrneC. 13.'. VCrerrt, 1' r aluiirg toclrniquev lii C Lii (t.Lt9 ,lec:n,'m13cnr4 cites Spec. Pabi. Ne. I Ar Irimici ii Sr 'ii V ''1 !clrthvli,varmd Iin"rlrcIll4',-Slor. It. 12.. Cd.; 'thur l'reec. mn 1 Zarhurr'ncnkr, T. Tnaxii' crqiiivalermcv fuelers ,i'!3s; fir Pnl!ls. Pn:tIDe, Pt tIFi fur ituuri;nmrr, trod wildlife. 1:'rr'rrnu. 11r'rltn t,awreimct', 13, 1311. '119: 'p :_.p It. SI. V'rr'brri fl (' ; all r.',ririairjir's jr s,,'kcy;'. 'juL. and cit_ia 'alumir. truer. 'tn:. Pr);....... 1910, 121, 751 770. Ea!omo/n, LI.. ('Jn'sr''ri, I.. S. Crcen)rcr9. .5. 0.. Fda. 9renn;n/nrnt Perspc'CL (9911, JOG. 775 r:.rlianparamive Ioieiry cl '.;F2'.il.ir'tr..,n'Iric'r.irlihemmzo.p_dicroimi (a sevL'rr frmnlr',vmtarlish '.pm''lr; m'Iurimmg orj'Ivlile' .a,nr developtrrea). mIth\ nan:iainua, cSriunaeicjrl fl',tr'r l'\'orkri aumocili' In, .ini] Waler' Crrvirnrrrrlerrt Fedcrn!iaim: IV'rchingiurl. I Ft. C., J':'15; Snrii,.i 21111). Incamrdona, J. P.; C liner T, K.; ScboIz, N, F. Def'r'a rn c.rrdiac furictutu prem'.nin' n;a'u-pinahrigv ahminrr'uulrne'a in air n'rrrbryra nrxpon.odlo pcnivcviir: iroirmalic hvnirecur'burra. Jan'.. "/'Vi' /'lnr ni. 2004, 195, lOt. 132. 1131 Eirrrica, El. F., Spn'bae. ii. I..; Fbolcowb, C. ; .. Johns' tm, 14. 11.; t'm'rma;nrndr'z, 3, Ft.; Ericksair , ii. 1.; l'isLge. 3. Ia; iook, P. IC. .\Jn'i!mrrn!t Jon 11:,' l,L',rnn;nnn,n;unr,rnf ll'crO'r url L1)nrn';r'nr:c'. lilir rrmt;Jmrmiem'icarr ISiErimo ti',ciilnt .)h.'ia.-p.lu,;iii "ii' J)r.nriLrcIm)9 anile Ii Fr''t.ige ru's tn; ta hr er lab 'c,' rica I i(nenr'nar'iri'mnn'Irro i,;n4Sni, 'l'rma'iarL. (991, 21, 2)0233, 1631 labeL 1;. '.1'.; Cauk, I' SI,; Pn'lv,rcjmm, F. P. 'lathe eJirlr;iln'iln'v fa:im'.rnol p tvchrIarilla lad drlreuzer.p.diaxirr dilrerrerafurari, and .tejun,'ri. !rrrMrn:n,n. l'na,n'in'nri thins'. 1)11111. 17n3i, .173 .51; liii Cunk,t'.M.; ti.rrbbirrn.l. A,; EmtLIrcaiI, Ft. I"n..thdpca.K.PnaGuarrey. P. IN; Walker, 13. K,: libel, 11.10; i'n'lersn nfl, Il. 1). Elfecisofarvi Irvdrncarbrar recn'pr'at .nrteii,ni ed n'uela'iitc singe nariril omr luke Oct11 popLiI;ntiauts rim Ink,' m,niirLrj,m ijir) rg rIm' 2tith I'r'nrnrnry. Eor,'e'n,n;, ('1. 'I'ncl.'n.'s'I. 2111,1:1. .17, 11.5. hnvirurrrrriyrn I.rl I 'FOrm' (Intl ta 'I rev 5:rlLmnn.nj'nmn.nL,vjncfaf 107,1 Pika, .5. 15'.: Eta. St. 1). 'Ph' learn' rc'r.Jn:nt'.e nI' 931w çnercin. Wuier Enrn/n/r m rnr'j'inn.(,'ernva'an,'. EPA 122-Z-'i'i.rliJl Mjm'er, F. F. 1.; l,1hli'riia'k. 51. It. :'nlinnl urn! i.! era I' :.x.'rcnrr" !anrer;mrm'tnili,'n,r i,'i;il ,la;nnlmran' it' .1111 ,'/:rarrinn nOn trim! in. rpia'r'a of Jn'n'nnn JJmn:''na'm'inn .tfi,,'i;m;/ ia irrlecii 'Lit WI Ill ire' tnrel,mrerr'.mrirt ''Vol m,I,'lm'9nitnn?uji l,r'r'mqi flntirau, 'ill. i":'n,''nnai1 0r'n' (9101, fritn)mm.elmm'rcrrmimnnrb'. )in'.''nnrrui Puirli-;rnimm iii. GO; 1.1_S. I.lipart. ''1 (00,1 Tavlrr,I..!i.;l)ntI,.11. K,;tmlivaki' 'I' ;5.ime.I K i:';'.'p;r''ssicic.. i,,'lcta;n,'d ''lii; rrneracnn'nc',ir'1;jm'c'l'.-10,m,4,'bmnllnnni,'ri'r'iu I tt'r'irnIrdal rmme'rmt of the Iii),....nat-, Flair arid 5'. i].dIrl'e' Sn'vies 5t'aahirrgtu'ir. ijiye!i'.t:'p.'rr'h, 1;.n'."a/JrrEa'.'mr2 l''i,',isln'ii dn'velcpmrreitn and DC. 113116. in)r'LLLIir'rU. ii iii bartn: amid rlrn'itrdrnnid 1' .'u' ,'Intcrn, flit: iris.!, I 'Cl I, 1011. 2)1'' H (513) Georgitcukia. F.; KIrnrr, tnt A. P.). i'nxin'itvn,iltlre pbutisinrtreri1t cyelodiemne imrs,vrirides la fresImwrirer ;rnirraai. C A. 11th! Swc'e'r.4mutd. Sn;L Soc. 1!, 1971. 233. 120 31. (57) Citrus'. 1. P.; Soioruonr, K. IF; Finals, 1. F.; Drort, K'. P.: r..i.:idLmmn. I. IC.; Ketiagu., to. to. ChlorpSTiten; ecriogi lii risk an',m'si a 'iii in Niarl'hhurerirsmen aquatic encirommracrri 101: l:it,'inii. r:,orrnnn,r, Tuxir&, 1' ..0. 160, 1" 1 29. (SlIt Sammdahj. 3, F.; Jrnktua, 1. 1. Pacrfnc Ce,ICULerd :rn,;n'n,n1ri'ir'luo i'!r'e) expomed to cirirnrpvrifns: hncinrrr,urk c'lnm:m'uir,ttt Ft eullinateu fnrr ucctv]cltoiirrenteruse itihiLiirietr. Fm:n'Lnannr 'ía: Chew, '2002,21, 2.1.32 2.150. (59) Harris, C. I..: Sir c,rb''. '1'. 1,.,; sl'rcali'', 1", 1).; Hur'stis, .5. 5'. ltm':ubm'vmtnjXjn'iIt- at 'x;r,'jcrn lOin like Oiti,'tr'ir r,nirmbcrw trout (L9rn'or)nv,rn'L'mna nrrn'i'/ari I.'. i.:ipmin i nrr:d.rk;n inlOr ltutrr'o, B. l'rtlrialngicab. c1rungc in m'vprurrLnt (i'' irrI't'ctn'd be B:ncn'phrri;ia /na fsmencrrp)trna 1h'nm'r, 11127 mid 0) ,'n;r'n'N iii,' ml i'ZIr'glnt. I traCt nhr'inr:errarianrr (l'remrrniod;n, Brmcn'pnImilidim'i..'lcini t9nti, 27, 24t -24G. jmk to parasites grows srroergcr. .t'rc'rt.'n' ('199, 24. itni'La,'Z,n.1 151.'. Till Kirai'r, I. I 2(5! '7) n Lij'nhurrs. V.;Llhaaarr.s,C, Srraiiciartb'cnis("rIrl;''H'oe rer'irt.rti,rrof ,,,'liiai':'a LctI;msiS trstrlsLml'tssic'nni. Pm'nn'ciailo!. Trndnny IlIrlir, 413 3 17. 73! Kit'i.''k,i'r, 1. .13. svmrergoettm betweirrt n'irmtn;nmade ijrk''micrii arm] m'stLcinCm'xpauei': Slink to arrrphi ni nil ilr'li,j'jrrmlrn's in rralure, lOan' torn;! ,knc( Sn'; USA. 2(102, 11.9, 0000 9004. irio.'.'rs Inrarnn'r). .tirrtcrear. 'l'nnancol. r trait, I I Pr, I'i°'i I lint (00) Viltalobus, S. A,; Papeanilias. Ft. SI ; Parer. S In,', ti.Ianikertship. A. F.; Meadows, 0; Tilti 11. 11. F.; I icr, I. F, 'tOxic lv 01' o,)1.lSDhi to mmrdakam d.rI( strain after n rim".. Mine enimhrrnrr;n' rrpsiiru' b 3OEn. ENVIRON?.IEt.ITa,L SCIENCE & IECHNOLOGe VOL. 35 N'). 10, 200E 0n''iista(inlu' ,'';';,';l Sn'ltC'nr:inr'i' 1,), 21,1,1), ,E.a'i.cm'n'! n/Lnu;l.ec'i'rJ)l .1i.rrn3r 11. 21)12 Ac'c'i'tn'i/.5U,i'!i 17, 2ofkS. (;5ii .1)157111